Behavioral and Biochemical Defects in Temperature-Sensitive Acetylcholinesterase Mutants of DROSOPHILA MELANOGASTER

Temperature-sensitive (ts) mutants of the Ace gene, which codes for acetylcholinesterase (AChE) in Drosophila melanogaster, were analyzed for defects in viability, behavior and function of the enzyme. The use of heat-sensitive and cold-sensitive mutations permited the function of AChE in the nervous system to be analyzed temporally. All ts mutations were lethal, or nearly so, when animals expressing them were subjected to restrictive temperatures during late embryonic and very early larval stages. Heat treatments to Ace-ts mid- and late larvae had little effect on the behavior of these animals or on the viability or behavior of the eventual adults. Heat-sensitive mutants exposed to nonpermissive temperatures as pupae, by contrast, had severe defects in phototaxis and locomotor activity as adults. AChE extracted from adult ts mutants that had developed at a permissive temperature were abnormally heat labile, and they had reduced substrate affinity when assayed at restrictive temperatures. However, enzyme activity did not decline during exposure of heat-sensitive adults to high temperatures even though such treatments caused decrements in phototaxis (29°) and, eventually, cessation of movement (31°). The cold-sensitive mutant also produced readily detectable levels of AChE when exposed to a restrictive temperature during the early developmental stage when this mutation causes almost complete lethality. We suggest that the relationship among the genetic, biochemical and neurobiological defects in these mutants may involve more than merely temperature-sensitive catalytic functions.     

Genetic,cytological, and ultrastructural characterization of a temperature-sensitive lethal in Drosophila melanogaster

Genetic analysis of a strain of Drosophila melanogaster revealed that a recessive mutation [l(1)ER^ts] causing temperature-sensitive embryonic lethality is located in the distal region of the X chromosome approximately at map position 18. At 22–25°C mutant embryos exhibit normal viability, and all eggs arrest prior to gastrulation if they are reared at 29°C. The mutant is biphasic, exhibiting a maternal effect which is expressed throughout the first 8 hr of development as well as a second temperature-sensitive period (TSP) during the first 3 days of larval life. Larvae exposed to the restrictive temperature (RT) during the second TSP must also spend the remainder of larval and pupal life and the time of normal eclosion at RT to die as fully developed pupae which fail to eclose. Light and electron microscopy of arrested embryos reveal disturbances in the distribution of nuclei, cytoplasm, and yolk and abnormal configurations of rough endoplasmic reticulum. The cause of pupal death during the second lethal period is unknown.     

Temperature sensitivity of muscle and neuron differentiation in embryonic cell cultures from the Drosophila mutant, shibire.

The sex-linked temperature-sensitive mutation, shibire^ts1, which causes, at the restrictive temperature, adult paralysis and pleiotropic morphological defects in embryonic, larval, and pupal development, has been shown to exhibit temperature-sensitive inhibition of differentiation in embryonic cultures in vitro. When shi cultures were incubated at 30°C for 24 hr, both muscle and neuron differentiation were inhibited more than 90% compared to control shi cultures incubated at 20°C. Heat shift experiments showed that the temperature-sensitive periods for neuron and muscle differentiation occurred at 11 to 18 and 14 to 16 hr, respectively, where zero time was the initiation of gastrulation in donor embryos. Short heat pulses (4 and 8 hr) which extended into the temperature-sensitive period resulted in moderate inhibition of differentiation; greater inhibition occurred as the duration of the pulses increased. In contrast, heating wild-type Oregon-R cultures at 30°C for 24 hr did not inhibit muscle cell differentiation and inhibited neuron differentiation relatively little. The temperature-sensitive period in shibire for muscle differentiation occurred well after myoblast division, during the period of myocyte elongation, aggregation, and fusion, whereas that for neuron differentiation took place during a period of enzyme synthesis (acetylcholinesterase and choline acetyltransferase) and axon elongation. Thus, the shi temperature-sensitive gene product affects at least two different cell types, in vitro, at different times during differentiation.     

Single Site Suppressors of a Fission Yeast Temperature-Sensitive Mutant in cdc48 Identified by Whole Genome Sequencing

The protein called p97 in mammals and Cdc48 in budding and fission yeast is a homo-hexameric, ring-shaped, ubiquitin-dependent ATPase complex involved in a range of cellular functions, including protein degradation, vesicle fusion, DNA repair, and cell division. The cdc48⁺ gene is essential for viability in fission yeast, and point mutations in the human orthologue have been linked to disease. To analyze the function of p97/Cdc48 further, we performed a screen for cold-sensitive suppressors of the temperature-sensitive cdc48-353 fission yeast strain. In total, 29 independent pseudo revertants that had lost the temperature-sensitive growth defect of the cdc48-353 strain were isolated. Of these, 28 had instead acquired a cold-sensitive phenotype. Since the suppressors were all spontaneous mutants, and not the result of mutagenesis induced by chemicals or UV irradiation, we reasoned that the genome sequences of the 29 independent cdc48-353 suppressors were most likely identical with the exception of the acquired suppressor mutations. This prompted us to test if a whole genome sequencing approach would allow us to map the mutations. Indeed genome sequencing unambiguously revealed that the cold-sensitive suppressors were all second site intragenic cdc48 mutants. Projecting these onto the Cdc48 structure revealed that while the original temperature-sensitive G338D mutation is positioned near the central pore in the hexameric ring, the suppressor mutations locate to subunit-subunit and inter-domain boundaries. This suggests that Cdc48-353 is structurally compromized at the restrictive temperature, but re-established in the suppressor mutants. The last suppressor was an extragenic frame shift mutation in the ufd1 gene, which encodes a known Cdc48 co-factor. In conclusion, we show, using a novel whole genome sequencing approach, that Cdc48-353 is structurally compromized at the restrictive temperature, but stabilized in the suppressors.     

Temperature-sensitive periods and autonomy of pleiotropic effects of l(1)Nts1, a conditional notch lethal in Drosophila.

Analysis of temperature-shift experiments using strains homo- and/or hemizygous for a temperature-sensitive (ts) mutation of the Notch locus, l(1)N^ts1, has permitted us to localize temperature-sensitive periods (TSPs) both for lethality and for adult ectodermal morphology defects. Discrete TSPs for lethality are localized to the first half of the embryonic period, to the second larval instar, to the third larval instar, and to a 15 hr period immediately after pupation. TSPs for adult morphology defects are localized to the second and third larval instars for eyeless-headless and duplicated antenna, to the third larval instar for small and rough (spl-like) eye, eye scar, fused leg segments, shortened tarsal leg segments, Notch wings, and extra macrochaetae, and to the early pupal period for extra and missing microchaetae, fa^g-like rough eye and thick wing vein defects. Within the third larval instar, distinct patterns of eye, wing, and leg defects are observed. There is a striking similarity between the adult morphology defects and TSPs of l(1)N^ts1 and those of the larval and adult locomotor mutant, shi^ts1 (C. A. Poodry, L. Hall, and D. T. Suzuki, 1973, Develop. Biol.32, 373–386). Expression of l(1)N^ts1 also has been studied in genetic mosaics, in which we find that the pleiotropic effects of l(1)N^ts1 are autonomously expressed.     

Studies of l(3)c43hs1 a polyphasic, temperature-sensitive mutant of Drosophila melanogaster with a variety of imaginal disc defects.

The heat-sensitive, lethal mutation l(3)c43^hs1 (3–49.0) produces wide variety of defects in the imaginal discs of Drosophila melanogaster. At permissive temperatures (20°C or lower), homozygotes are viable, but sterile. At 22°C, lethality occurs during the late pupal stage, and at 25°C or higher, lethality occurs during the third larval instar. The imaginal-disc abnormalities observed after exposure to restrictive temperatures include: deficiencies of head structures, duplications and deficiencies of the antenna, a homeotic transformation of the arista to tarsus, duplications and deficiencies of wing and haltere structures, differentiation of amorphous cuticular material in the wing blade, an increase in the number of sex-comb teeth, and disruption of the normal segmentation of the tarsus. Exposure to 27°C for 24 hr at different times in the life cycle revealed that each of these defects has a characteristic temperature-sensitive period (TSP) during the larval stages. Injection of wing discs before and after their TSP showed that the mutation is expressed autonomously. These results are discussed in relation to the role that the l(3)c43⁺ gene plays in the development of imaginal discs.     

Expression of phenotypes in a temperature-sensitive allele of the apterous mutation in Drosophila melanogaster

A temperature-sensitive allele of the apterous (ap) locus of Drosophila melanogaster has been used to examine the phenotypes produced by this mutation, which include wing, mesonotal, and haltere deficiencies, precocious adult death, and nonvitellogenic oocyte development. When raised at 15°C, homozygous ap^ts78j adults have nearly wild-type wing morphology except for patches of missing triple-row bristles and posterior wing margin deficiencies. With the exception of two missing bristles, the dorsal mesonotum and the haltere appear as wild-type. Increasing deficiency of structures derived from the wing and haltere imaginal discs results from increasing culture temperature, and at 29°C, the wing blade, many dorsal mesonotal bristles, and the haltere are absent. The temperature-sensitive period in development for these deficient phenotypes extends from late-second to mid-third instar. Despite extensive deficiencies seen after ap^ts78j larvae are heat-pulsed at 29°C, no duplication of the notal structures is evident, a common response of other mutants having extensive wing deficiencies. When raised at 29 or 25°C, ap^ts78j adults are short-lived, and females show nonvitellogenic oocyte development. At 22°C, however, adults are long-lived, and females are vitellogenic and lay fertile eggs. A sharp temperature-sensitive period for both phenotypes is located during the first 24 hr of pupal development. The application of a juvenile hormone mimic, ZR-515, restored vitellogenesis to ap^ts78j females raised at 25°C but was unable to rescue them from precocious death.     

The Drosophila Tumorous lethal hematopoietic oncogene is a dominant mutation in the hopscotch locus

The Drosophila Tumorous-lethal (Tum-l) mutation acts as an activated oncogene, causing hematopoietic neoplasms, overproliferation, and premature differentiation. Tum-l is a dominant mutation in the hopscotch (hop) locus, which is required for cell division and for proper embryonic segmentation. The Tum-l temperature-sensitive period for melanotic tumor formation includes most of larval and pupal development.     

Alterations in the production of hemocytes due to a neoplastic mutation of Drosophila melanogaster

The hemocytes of a genetically induced, temperature-sensitive lethal mutation of Drosophila, Tum¹, were examined both quantitatively and qualitatively during the third larval instar. At the tumor-permissive temperature, 29°C, there was a fourfold increase in the concentration of circulating hemocytes in mutant larvae as compared to control. Additionally, the relative frequency of lamellocytes was 30 times greater in Tum¹ larvae than Basc in the early third instar. However, the severity of this abnormality gradually diminished as Tum¹ approached pupariation; though high frequencies of lamellocytes were always present. At the tumor-restrictive temperature (15°C) the concentration of circulating hemocytes was over twice that found at 29°C for Tum¹ larvae, and did not change during the course of third instar. However, in contrast to 29°C there was no abnormal increase in the frequency of lamellocytes at the tumor-restrictive temperature. Control larvae had equivalent concentrations of hemocytes at both temperatures. In one of two temperature shift experiments, Tum¹ larvae shifted from 15° to 29°C at the beginning of third instar expressed the abnormal hemocyte concentration and differentiation associated with larvae raised continuously at 29°C. In addition, Tum¹ larvae shifted from 29° to 15°C expressed reduced abnormalities of hemocyte differentiation, e.g., with fewer lamellocytes in circulation. The possibility of a temperature-sensitive period for the activation of the Tum¹ gene is discussed.     

Developmental properties of Shibire: a pleiotropic mutation affecting larval and adult locomotion and development

The sex-linked recessive mutation, shibire^ts1, causes rapid paralysis in adults and larvae at 29°C whereas normal mobility occurs at 22°C. The influence of the mutation on development was analyzed by studying the effects of brief heat pulses administered at various developmental stages. Mutant animals had a polyphasic temperature-sensitive period (TSP) for lethality and a polyphasic effective lethal phase (LP). In addition, heat shocks caused a broad spectrum of phenotypically distinct defects, each characteristic having its own distinct TSP. Some of the processes affected were: early embryogenesis including gastrulation, and the development of eyes, bristles, legs, wings, and the neuromuscular system. The developmental properties of shibire^ts1 allowed us to make a number of observations concerning determination and pattern formation and to conclude that the lesion affects a fundamental cell process common to many cell types.     

Cold-Sensitive Cell-Division-Cycle Mutants of Yeast: Isolation, Properties, and Pseudoreversion Studies

We isolated 18 independent recessive cold-sensitive cell-division-cycle (cdc) mutants of Saccharomyces cerevisiae, in nine complementation groups. Terminal phenotypes exhibited include medial nuclear division, cytokinesis, and a previously undescribed terminal phenotype consisting of cells with a single small bud and an undivided nucleus. Four of the cold-sensitive mutants proved to be alleles of CDC11, while the remaining mutants defined at least six new cell-division-cycle genes: CDC44, CDC45, CDC48, CDC49, CDC50 and CDC51.—Spontaneous revertants from cold-sensitivity of four of the medial nuclear division cs cdc mutants were screened for simultaneous acquisition of a temperature-sensitive phenotype. The temperature-sensitive revertants of four different cs cdc mutants carried single new mutations, called Sup/Ts to denote their dual phenotype: suppression of the cold-sensitivity and concomitant conditional lethality at 37°. Many of the Sup/Ts mutations exhibited a cell-division-cycle terminal phenotype at the high temperature, and they defined two new cdc genes (CDC46 and CDC47). Two cold-sensitive medial nuclear division cdc mutants representing two different cdc genes were suppressed by different Sup/Ts alleles of another gene which also bears a medial nuclear division function (CDC46). In addition, the cold-sensitive medial nuclear division cdc mutant csH80 was suppressed by a Sup/Ts mutation yielding an unbudded terminal phenotype with an undivided nucleus at the high temperature. This mutation was an allele of CDC32. These results suggest a pattern of interaction among cdc gene products and indicate that cdc gene proteins might act in the cell cycle as complex specific functional assemblies.     

Genetic interaction of DED1encoding a putative ATP-dependent RNA helicase with SRM1 encoding a mammalian RCC1 homolog in Saccharomyces cerevisiae

The Saccharomyces cerevisiae temperature-sensitive mutants srm1-1, mtr1-2 and prp20-1 carry alleles of a gene encoding a homolog of mammalian RCC1. In order to identify a protein interacting with RCC1, a series of suppressors of the srm1-1 mutation were isolated as cold-sensitive mutants and one of the mutants, designated ded1-21, was found to be defective in the DED1 gene. The double mutant, srm1-1 ded1-21, could grow at 35°?C, but not at 37°?C. A revertant of srm1-1 ded1-21 that became able to grow at 37°?C acquired another mutation in the SRM1 gene, indicating the tight relationship between SRM1 and DED1. In all the rcc1 ^- strains examined, the amount of mutated SRM1 proteins was reduced or not detectable at the nonpermissive temperature. While mutated SRM1 protein was stabilized in all of the rcc1 ^- strains by the ded1-21 mutation, the ded1-21 mutation suppressed both srm1-1 and mtr1-2, but not the prp20-1 mutation, contrary to the previous finding that overproduction of the S. cerevisiae Ran homolog GSP1 suppresses prp20-1, but not srm1-1 or mtr1-2.     

Mutations in gfpt1 and skiv2l2 cause distinct stage-specific defects in larval melanocyte regeneration in zebrafish

The establishment of a single cell type regeneration paradigm in the zebrafish provides an opportunity to investigate the genetic mechanisms specific to regeneration processes. We previously demonstrated that regeneration melanocytes arise from cell division of the otherwise quiescent melanocyte precursors following larval melanocyte ablation with a small molecule, MoTP. The ease of ablating melanocytes by MoTP allows us to conduct a forward genetic screen for mechanisms specific to regeneration from such precursors or stem cells. Here, we reported the identification of two mutants, eartha^j23e1 and julie^j24e1 from a melanocyte ablation screen. Both mutants develop normal larval melanocytes, but upon melanocyte ablation, each mutation results in a distinct stage-specific defect in melanocyte regeneration. Positional cloning reveals that the eartha^j23e1 mutation is a nonsense mutation in gfpt1 (glutamine:fructose-6-phosphate aminotransferase 1), the rate-limiting enzyme in glucosamine-6-phosphate biosynthesis. Our analyses reveal that a mutation in gfpt1 specifically affects melanocyte differentiation (marked by melanin production) at a late stage during regeneration and that gfpt1 acts cell autonomously in melanocytes to promote ontogenetic melanocyte darkening. We identified that the julie^j24e1 mutation is a splice-site mutation in skiv2l2 (superkiller viralicidic activity 2-like 2), a predicted DEAD-box RNA helicase. Our in situ analysis reveals that the mutation in skiv2l2 causes defects in cell proliferation, suggesting that skiv2l2 plays a role in regulating melanoblast proliferation during early stages of melanocyte regeneration. This finding is consistent with previously described role for cell division during larval melanocyte regeneration. The analyses of these mutants reveal their stage-specific roles in melanocyte regeneration. Interestingly, these mutants identify regeneration-specific functions not only in early stages of the regeneration process, but also in late stages of differentiation of the regenerating melanocyte. We suggest that mechanisms of regeneration identified in this mutant screen may reveal fundamental differences between the mechanisms that establish differentiated cells during embryogenesis, and those involved in larval or adult growth.     

Rhythmic Leaflet Movement in Albizzia julibrissin: Effect of Electrolytes and Temperature Alteration

The rhythmic movement of darkened Albizzia leaflets is accompanied by K⁺ flux in pulvinule motor cells whose turgor changes control opening and closing. The azide-sensitive open phase is promoted by an increase in temperature from 16 to 33C (Q₁₀ = 3), implying active transport of K⁺ ions during this period. The azide-insensitive closed phase is less temperature-sensitive and has a Q₁₀ less than 1, implying diffusion or some other physical process as the predominant pathway of K⁺ flux at this time. Thus rhythmic leaflet movement is probably due to oscillation in active K⁺ transport or membrane permeability or both. External electrolytes (0. 1 n) alter leaflet angle during the open, but not the closed, phase of the rhythm. All chlorides except NH₄⁺ promote opening, with divalent more effective than monovalent ions. Some anions promote and others inhibit opening; activity is not correlated with charge. It is likely that electrolytes alter leaflet movement by altering K⁺ flux, accomplishing this by interacting with key macromolecules in motor cell membranes.     

Genetic control of DNA initiation in Escherichia coli

We describe the isolation, and properties of a mutant (CT28) of Escherichia coli with a temperature-sensitive defect in DNA initiation that is reversible. The mutation (dna-28) responsible for this defect is shown to be located in the same region of the map as the dnaC group of DNA initiation mutants.A terminalized culture of CT28 initiates DNA synthesis synchronously immediately upon lowering the temperature, and will do so in the presence of chloram-phenicol.During prolonged incubation at the non-permissive temperature, the cells accumulate a capacity to initiate multiple rounds of replication per chromosome.The variation in the susceptibility of the argH^? and thyA^? alleles to reversion by pulse mutagenesis with nitrosoguanidine during a synchronous round of DNA replication, suggests that this round of replication is bidirectional and commences from an origin in the vicinity of 60 to 65 minutes.CT28 contains two temperature-sensitive mutations. These have been mapped and separated into two derivative strains. One of these, CT28-3b, carries the dna-28 mutation of the C locus, and like the parental double mutant is reversibly temperature-sensitive for an initiation function; but it is more temperature-sensitive than either the double mutant or the other single mutant derivative, CT28-1. The other, CT28-1, is not defective in DNA replication or initiation of replication at the non-permissive temperature.     

Hyperthermia of male <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> meiocytes induces abnormalities in both paternal and maternal sex chromosome sets of the offspring

Nondisjunction and loss of sex chromosomes caused by exposure of male Drosophila melanogaster to heat shock (HS) (37°C for 1 h) has been studied to determine the role of mutation l(1)ts403 (sbr ¹⁰) in the control of chromosome segregation during cell division. Hyperthermia of males at the pupal stage has been demonstrated to increase the number of offspring with abnormalities of not only paternal, but also maternal sex chromosome sets. According to the criterion used, there is a temperature-sensitive period of spermatogenesis, which presumably coincides with meiosis. Phenotypes of some individuals correspond to the presence of two sex chromosomes of obtained from the same parent. The frequency of abnormal chromosome sets in the off-spring of male carriers of the sbr ¹⁰ mutation is about two times higher than in the offspring of males without this mutation.     

Developmental timing of a sensory-mediated larval surfacing behavior correlates with cessation of feeding and determination of final adult size

Controlled organismal growth to an appropriate adult size requires a regulated balance between nutrient resources, feeding behavior and growth rate. Defects can result in decreased survival and/or reproductive capability. Since Drosophila adults do not grow larger after eclosion, timing of feeding cessation during the third and final larval instar is critical to final size. We demonstrate that larval food exit is preceded by a period of increased larval surfacing behavior termed the Intermediate Surfacing Transition (IST) that correlates with the end of larval feeding. This behavioral transition occurred during the larval Terminal Growth Period (TGP), a period of constant feeding and exponential growth of the animal. IST behavior was dependent upon function of a subset of peripheral sensory neurons expressing the Degenerin/Epithelial sodium channel (DEG/ENaC) subunit, Pickpocket1(PPK1). PPK1 neuron inactivation or loss of PPK1 function caused an absence of IST behavior. Transgenic PPK1 neuron hyperactivation caused premature IST behavior with no significant change in timing of larval food exit resulting in decreased final adult size. These results suggest a peripheral sensory mechanism functioning to alter the relationship between the animal and its environment thereby contributing to the length of the larval TGP and determination of final adult size.     

Developmental genetics of a temperature-sensitive RNA polymerase II mutation in Drosophila melanogaster

Summary Purified RNA polymerase II (RNA nucleotidyl-transferase; EC 2.7.7.6) extracted from flies possessing lesions in the Ultrabithorax-like (Ubl) locus of Drosophila melanogaster has altered activity in vitro (Greenleaf et al. 1979, 1980; Coulter and Greenleaf 1982). This strongly suggests that the Ubl locus encodes a subunit of RNA polymerase II. Ethyl methanesulfonate was used to induce a temperature-sensitive mutation in this locus. Flies either homozygous or hemizygous for this new X–linked mutation (Ubl ^ts) display viability comparable to that of wild-type flies at 22° C but are lethal at 29° C. The temperature-sensitive period for Ubl ^ts flies is between gastrulation (6 h, 29° C) and pupation (9–10 days, 22° C). Zygotes shifted from 22° C to 29° C die at either the late embryonic or first larval instar stage while temperature shifts of second and third instar larvae result in the lethal phase occurring at the pupal stage. Most pupae shifted from 22° C to 29° C undergo metamorphosis and eclose as adults. Adults are viable if placed at 29° C; however, all females and some males become sterile if maintained at this temperature.Somatic recombination was used to induce clones homozygous for a null allele of Ubl at different stages of development. Clones of this null allele appear to be cell lethal indicating that the Ubl ⁺ gene product is required at all stages of development. The viability of Ubl ^ts pupae and adults at 29° C may result from only a partial reduction in activity caused by the mutation at this nonpermissive temperature.     

Analysis of molting and metamorphosis in the ecdysteroid-deficient mutant L(3)3DTS of Drosophila melanogaster

The dominant temperature-sensitive mutation L(3)3^DTS (DTS-3) in Drosophila melanogaster causes lethality of heterozygotes during the third larval instar at the restrictive temperature (29°C). Temperature-shift experiments revealed two distinct temperature-sensitive periods, with lethal phases during the third larval instar (which may persist for 4 weeks) and during the late pupal stage. At 29°C mutant imaginal discs are unable to evert in situ, but did evert normally if cultured in the presence of exogenous ecdysterone or when implanted into wild-type larval hosts. The only morphologically abnormal tissue present in the lethal larvae is the ring gland, the prothoracic gland being greatly hypertrophied in third instar DTS-3 larvae. Injection of a single wild-type ring gland rescued these mutant larvae, indicating that the mutant gland is functionally, as well as morphologically, abnormal. Finally, the mutant larvae were shown to have less than 10% of the wild-type ecdysteroid levels. These results are all consistent with a proposed lesion in ecdysteroid hormone production in DTS-3 larvae. A comparison with the phenotypes of other “ecdysone-less” mutants is presented.     

A model investigation of the impact of ventilation-perfusion mismatch on oxygenation during apnea in preterm infants

Ventilation-perfusion (V/Q) mismatch is a prominent feature of preterm infants and adults with lung disease. V/Q mismatch is known to cause arterial hypoxemia under steady-state conditions, and has been proposed as the cause of rapid arterial oxygen desaturation during apnea. However, there is little evidence to support a role for V/Q mismatch in the dynamic changes in arterial oxygenation that occur during apnea. Using a mathematical model, we quantified the effect of V/Q mismatch on the rate of desaturation during apnea to ascertain whether it could lead to rates of up to 10% s^-1 as observed in preterm infants. We used a lung-body model for the preterm infant that incorporated 50 parallel alveolar-capillary units that were ventilated and perfused with the severity of V/Q mismatch (σ) defined conventionally according to σ=S.D. of the distribution of V/Q ratios. Average desaturation rate 10 s from apnea onset was strongly elevated with worsening V/Q mismatch as a result of an earlier desaturation of low V/Q units compared with high V/Q units. However, V/Q mismatch had little impact after apnea onset, with peak desaturation rate only substantially increased if mismatching caused a lowered resting arterial O₂ saturation. In conclusion, V/Q mismatch causes a more immediate onset of desaturation during apnea, and therefore places preterm infants and adults with lung disease at risk of hypoxemic dips. However, V/Q mismatch does not accelerate desaturation rate beyond apnea onset and cannot, therefore, explain the rapid desaturation observed during recurrent apnea in preterm infants.