Kidney function in the Spinifex hopping mouse, Notomys alexis

Notomys alexis (Spinifex hopping mouse) is found in the arid zone of Australia. The structure and function of the kidneys allow this species to conserve water. This study investigated the rate at which N. alexis can reduce urine volume and increase the concentration of electrolytes and solutes when water deprived. It also looked at the response to rehydration, following a period of water deprivation. The laboratory mouse, Mus musculus domesticus, was used for comparison. N. alexis is able to reduce urine volume and increase urine concentration more rapidly than M. m. domesticus when water deprived. This appears to occur prior to any measurable changes in plasma electrolyte concentrations and is not due to reductions in glomerular filtration rate. Gradual water deprivation over a period of 10 days allowed N. alexis to adjust so that urine composition was similar in many ways to animals that had ad libitum access to water, whereas M. m. domesticus required significant water supplementation to maintain body weight at 85% of initial body weight. Ability to concentrate urine rapidly is characteristic of a well-insulated renal medulla [Bankir, L., DeRouffignac, C., 1985. Urinary concentrating ability: insights from comparative anatomy. Am. J. Physiol. 249, R643-666]. However, a well-insulated medulla is normally associated with slow dilution of urine when animals are rehydrated. N. alexis was able to produce dilute urine very rapidly following rehydration of water deprived animals. Physiological control of renal function appears to be complex. Although M. m. domesticus is able to produce concentrated urine, it is unable to survive without free water and responds more slowly to water deprivation.     

Mating behaviour and insemination in the hopping mouse (Notomys alexis)

Hopping mice have extremely small testes and a reduced complement of male accessory sex glands. Nevertheless, greater than 1000 spermatozoa populate the isthmus of the oviduct after each insemination and a range of morphological types is found similar to that in the male tract. When females are primed with gonadotrophins they sometimes lock with more than one of the males within the group and in the present study two inseminations sometimes took place at the one oestrus. There is thus the possibility for intermale sperm competition within the female tract of these animals.     

Environmental factors and reproduction in the female hopping mouse, Notomys alexis.

Australian hopping mice (Notomy alexis) were subjected to short or long photoperiods, different social environments and/or water deprivation. The age at which vaginal opening and first oestrus occurred was delayed by short photoperiods and by the absence of male proximity. Water deprivation prevented normal maturation. The normal 7 to 8 day oestrous cycle was prolonged by short photoperiods or water deprivation. Animals deprived of water had lighter body, ovarian and uterine weights, and follicular growth seemed to be impaired. No differences in organ weights were found between animals in the two photoperiods. The significance of the findings is discussed.     

Quantitative estimation of islet tissue of pancreas in Spinifex hopping mouse (Notomys alexis)

Pancreases from three male and three female adult Spinifex hopping mice ( Notomys alexis ) were studied. No correlation was found between pancreas weight and body weight. Estimations of islet tissue mass and of individual cell types were made on paraffin sections of Bouin-fixed tissue taken from head, neck, body and tail regions of pancreas of each animal. Islet tissue mass was assessed using a linear scanning technique on sections stained with haematoxylin and eosin, and was compared with body weight. Specific cell types were assessed using a point-intercept method, on aldehyde-fuchsin-stained sections for beta (β) cells, and on immunoperoxidase labelled sections for alpha (α) cells (glucagon) and delta (δ) cells (somatostatin). Positive regional differences noted were a greater proportion of islet tissue in the tail region, and a lower proportion of α cells in the head region. Alpha cells were peripherally situated in the islets.
These results show some elements of agreement with a previously proposed hypothesis regarding the general patterns of arrangement of the mammalian endocrine pancreas.     

Organization of testicular interstitial tissue of an Australian rodent,the spinifex hopping mouse,Notomys alexis

Summary The organization of testicular interstitial tissue of the spinifex hopping mouse, Notomys alexis differs from that of other rodents. It comprises between 10.3% and 17.3% (average 15.0%) of the total testicular volume, and is variable in its organization both at different locations within the testis of the one animal and among different individuals. Abundant, closely packed Leydig cells are usually present; however, in some regions large, thick-walled blood vessels and extensive peritubular lymphatic spaces, often lacking an endothelium adjacent to the Leydig cells, are also prominent. The Leydig cells in contact with the large blood vessels and lymphatics, unlike those in regions where lymph is sparse, are not densely packed and sometimes contain numerous lipid droplets. Ultrastructure of Leydig cells is typical of steroid-producing cells; however, mitochondria are often extremely large, unusual in shape or bizarely arranged in relation to one another. Also electrondense bodies displaying a paracrystalline-like internal structure of parallel, electron-dense filaments arranged in a lattice pattern occur in the cytoplasm of many cells. The significance of these unusual ultrastructural features and the organization of the interstitial tissue remain to be determined conclusively, but may relate to steroid synthesis, secretion and uptake.     

Copulatory behaviour and coagulum formation in the female reproductive tract of the Australian hopping mouse, Notomys alexis

Hopping mice were examined to study two interrelated questions: (1) when groups of adults of both sexes are kept together in one cage in the laboratory is there evidence that the females copulate with only one male, and (2) is a copulatory plug formed in the female tract after ejaculation? The findings indicate that a female will sometimes lock with more than one male in the group during an oestrous period induced by administration of exogenous gonadotrophins, and that a small 'plug' of soft material forms post coitum in the more caudal parts of the female tract. Individuals of this species, therefore, do not appear to be strictly monogamous, at least in this artificial laboratory situation. Although a coagulum is formed, this is quite different from the typical hard copulatory plug that occurs in common laboratory murids; it may possibly reduce sperm backflow from the lower region of the female reproductive tract.     

Isolation of the mite Myocoptes musculinus Koch from the Spinifex Hopping mouse (Notomys alexis)

This paper reports on the isolation and identification of the fur-clasping mite, Myocoptes musculinus, from the faeces of the Spinifex Hopping mouse (Notomys alexis). This investigation adds to the sparse records of ectoparasites collected from native Australian murids.     

The effect of water deprivation on the expression of atrial natriuretic peptide and its receptors in the spinifex hopping mouse,Notomys alexis

This study investigated the mRNA expression of the atrial natriuretic peptide (ANP) system (peptide and receptors) during water deprivation in the spinifex hopping mouse, Notomys alexis, a native of central and western Australia that is well adapted to survive in arid environments. Initially, ANP, NPR-A and NPR-C cDNAs (partial for receptors) were cloned and sequenced, and were shown to have high homology with those of rat and mouse. Using a semi-quantitative multiplex PCR technique, the expression of cardiac ANP mRNA and renal ANP, NPR-A, and NPR-C mRNA was determined in 7- and 14-day water-deprived hopping mice, in parallel with control mice (access to water). The levels of ANP mRNA expression in the heart remained unchanged, but in the kidney ANP mRNA levels were increased in the 7-day water-deprived mice, and were significantly decreased in the 14-day water-deprived mice. NPR-A mRNA levels were significantly higher in 7-day water-deprived mice while no change for NPR-A mRNA expression was observed in 14-day water-deprived mice. No variation in NPR-C mRNA levels was observed. This study shows that water deprivation differentially affects the expression of the ANP system, and that renal ANP expression is more important than cardiac ANP in the physiological adjustment to water deprivation.     

The distribution of renal hyaluronan and the expression of hyaluronan synthases during water deprivation in the Spinifex hopping mouse, Notomys alexis

Hyaluronan (HA) is a glycosaminoglycan that is synthesized by a family of enzymes called hyaluronan synthases (HASs), of which there are three isoforms (HAS1, 2 and 3) in mammals. The HASs have different tissue expression patterns and function, indicating that synthesis of HA and formation of the HA matrix may be regulated by various factors. The HA matrix has an important role in renal water handling and the production of a concentrated urine. We investigated the distribution of HA and the expression of HAS1, HAS2 and HAS3 mRNAs in the kidney of the Spinifex hopping mouse, Notomys alexis, a native Australian desert rodent that is reported to produce the most concentrated urine of any mammal. After periods of three, seven and fourteen days of water deprivation, the distribution of renal HA changed considerably, and there was a general down-regulation of HAS mRNA expression. It is proposed that the regulation of HA synthesis by the different HAS isoforms during water deprivation in N. alexis, could be influenced by the molecular mass of the HA chains produced by each isoform, followed by the rate at which the individual HAS produces HA.     

The distribution of renal hyaluronan and the expression of hyaluronan synthases during water deprivation in the Spinifex hopping mouse, Notomys alexis.

Hyaluronan (HA) is a glycosaminoglycan that is synthesized by a family of enzymes called hyaluronan synthases (HASs), of which there are three isoforms (HAS1, 2 and 3) in mammals. The HASs have different tissue expression patterns and function, indicating that synthesis of HA and formation of the HA matrix may be regulated by various factors. The HA matrix has an important role in renal water handling and the production of a concentrated urine. We investigated the distribution of HA and the expression of HAS1, HAS2 and HAS3 mRNAs in the kidney of the Spinifex hopping mouse, Notomys alexis, a native Australian desert rodent that is reported to produce the most concentrated urine of any mammal. After periods of three, seven and fourteen days of water deprivation, the distribution of renal HA changed considerably, and there was a general down-regulation of HAS mRNA expression. It is proposed that the regulation of HA synthesis by the different HAS isoforms during water deprivation in N. alexis, could be influenced by the molecular mass of the HA chains produced by each isoform, followed by the rate at which the individual HAS produces HA.     

Regenerated luminal epithelial cells are derived from preexisting luminal epithelial cells in adult mouse prostate

Determining the source of regenerated luminal epithelial cells in the adult prostate during androgen deprivation and replacement will provide insights into the origin of prostate cancer cells and their fate during androgen deprivation therapy. Prostate stem cells in the epithelial layer have been suggested to give rise to luminal epithelium. However, the extent of stem cell participation to prostate regrowth is not clear. In this report, using prostate-specific antigen-CreER(T2)-based genetic lineage marking/tracing in mice, preexisting luminal epithelial cells were shown to be a source of regenerated luminal epithelial cells in the adult prostate. Prostatic luminal epithelial cells could survive androgen deprivation and were capable of proliferating upon androgen replacement. Prostate cancer cells, typically exhibiting a luminal epithelial phenotype, may retain this intrinsic capability to survive and regenerate in response to changes in androgen signaling, providing part of the mechanism for the ultimate failure of androgen deprivation therapy in prostate cancer.     

Collagen and the proliferation and differentiation of rat ventral prostate epithelial cells

The purpose of this study was to determine if a cause-and-effect relationship exists between androgen-induced changes in collagen and epithelial cell proliferation and/or differentiation in rat ventral prostate. Analyses of the temporal relationship between dihydrotestosterone (DHT)-induced changes in the synthesis and levels of collagen in the regressed ventral prostates of adult castrates demonstrated that, during the first 7 days of restoration of prostatic growth, androgen increased the synthesis as well as the degradation of collagen. Cis-hydroxyproline (CHP) treatment (2-200 mg/kg) during the first 7 days of androgen-stimulated prostatic growth, combined with maintenance of animals on a proline-free diet, produced a dose-dependent reduction in prostate weight and DNA content to a maximum of 50%. The epithelium was characterized by numerous disorganized layers of irregularly shaped and tightly packed cells, many of which had no contact with the basal lamina. There was a loss of epithelial lamina lucida and the development of a ragged lamina densa. Cis-hydroxyproline effects were reversible in that, following cessation of CHP treatment, the perturbed morphology, DNA content, and organ weight returned to the range of DHT-treated controls. Collagenous components seem to be important in supporting the normal androgen-dependent proliferation and differentiation of prostatic epithelial cells.     

Cytological organization of the seminiferous epithelium in the Australian rodents Pseudomys australis and Notomys alexis

Absolute and relative testis mass of two species of conilurine rodents, the plains rat (Pseudomys australis) and the hopping mouse (Notomys alexis), were markedly different. In plains rats the testis comprised approximately 3% of body mass while in hopping mice they rarely accounted for more than 0.2%. In both species, 8 cellular associations, or cycle stages, were recognized in the seminiferous epithelium. The relative frequencies, and hence durations, of Stages I to VIII were 5.1, 7.0, 12.5, 10.5, 5.4, 25.9, 4.4 and 29.0% of one cycle for plains rats (N = 14), and 20.4, 10.9, 9.6, 8.1, 5.1, 20.4, 14.7 and 10.8% for laboratory-bred hopping mice (N = 35). Generally, the relative durations of the cycle stages were very similar between plains rat individuals, but were variable between hopping mice. Also, organization of the seminiferous epithelium was less rigid in hopping mouse testes, and several anomalies were observed. These included: the occurrence of 2-4 cellular associations in about 20% (range 11.3-33.9%) of tubular cross-sections, deviations in cellular composition in certain cycle stages from that usually observed, and an increased incidence of degenerating cells, together with the presence of multinucleated "giant' cells, within the seminiferous epithelium.     

Polymorphism of the X-chromosome,Y-chromosome and autosomes in the Australian hopping mice,Notomys alexis,N. cervinus and N. fuscus (Rodentia,Muridae)

All three species of Notomys so far studied possess a diploid number of 48. Many elements in the karyotype of N. alexis are polymorphic due to variation in heterochromatin, but the variation is most marked in autosomal pair 1, which occurs in at least four forms, the X-chromosome, which occurs in three forms, and the Y-chromosome which occurs in many forms. N. cervinus is unique in the genus in possessing an entirely biarmed karyotype due mainly to the addition of heterochromatic short arms. The X-chromosome of N. cervinus occurs in three forms and the Y-chromosome in two forms. The karyotype of N. fuscus is mainly telocentric although two autosomal pairs are polymorphic due to pericentric inversions. The X- and Y-chromosomes both occur in two forms in N. fuscus. Chromosome measurements and C-banding show that most of the variation in the size and morphology of the sex-pair both within and between species is due to variation in constitutive heterochromatin.     

Hepatocyte growth factor supports androgen stimulation of growth of mouse ventral prostate epithelial cells in collagen gel matrix culture

To assess the role of hepatocyte growth factor (HGF) and androgen in growth of prostate epithelial cells, we isolated mouse ventral prostate epithelial cells and cultured them in a three-dimensional type I collagen gel matrix under serum-free conditions. Although the prostate epithelial cells tended to die in the insulin-supplemented basal medium, 5alpha-dihydrotestosterone (DHT) prevented the cell death, and HGF slightly stimulated the growth. By contrast, coexistence of DHT and HGF greatly augmented the growth and branching morphogenesis of the epithelial cells. Some of the outgrowths formed under these conditions showed enlarged structures resembling the prostate ducts or alveoli. Examination of the stromal cell-conditioned medium revealed that a growth-stimulating activity is present in the conditioned medium. A major portion of this activity was abolished by anti-HGF IgG. These observations suggest that HGF is produced by the stromal cells of the prostate gland and supports the androgen stimulation of growth of the epithelial cells.     

Vocalizations of Australian hopping mice (Rodentia: Notomys)

The four species of hopping mouse studied, N. alexis, N. cervinus, N. fuscus and N. mitchellii , were found to have a basic repertoire of eight more or less discrete vocalizations: three in the young—the calls produced in the nest, ultrasonic pipping given when removed from the nest and a high intensity (pain) squeal—and five in the adult—a low intensity pip given during mutual grooming, a high intensity (pain) squeal, a medium intensity squeak given when "annoyed", twittering given during aggressive chases and ultrasonic pipping given during non-aggressive encounters. The calls of the different species varied in structure and in pitch. No vocalizations were associated with threat or fighting. Calls given in the nest by suckling young are well developed and the intensity of such calls appears to mirror the general motivational state of the caller. It is suggested that these calls may act to maintain maternal behaviour in the mother.     

Receptors for vasoactive intestinal peptide on isolated epithelial cells of rat ventral prostate

Receptors for porcine vasoactive intestinal peptide have been characterized in isolated epithelial cells of rat ventral prostate. The interaction of ¹²⁵I-labelled VIP with cells was rapid, reversible, specific, saturable and dependent on temperature. Degradation of peptide and receptors was minimized at 15°C. At apparent equilibrium, the binding of ¹²⁵I-labelled peptide was competitively inhibited by native VIP in the 1·10⁻¹⁰−10⁻⁷ M range concentration. The binding data were compatible with the existence of two classes of receptors: a high-affinity class with a K_d = 4.0 nM and a low binding capacity (0.12 pmol VIP/mg cell protein), and a low-affinity class with a K_d = 17.8 nM and a high binding capacity (1.6 pmol VIP/mg cell protein). Chicken VIP and porcine secretin exhibited a 7-fold higher and a 7-fold lower affinity than porcine VIP for binding sites, respectively. Glucagon, Leu-enkephalin, Met-enkephalin and somatostatin were ineffective. The presence of high-affinity receptors for VIP together with previous reports on the occurrence of VIP-containing neurones innervating the male genitourinary tract strongly suggest that this peptide may be important in the physiological regulation of the functions of prostatic epithelium.