Immunohistochemical identification of substance P in cutaneous sensory organs (electroreceptors) of gymnotid teleost fish

Summary The distribution of the neuropeptide substance P, which is considered to be a neurotransmitter or neuromodulator of the central nervous system, has been studied in the cutaneous electroreceptor organs (tuberous and ampullary organs) of 3 species of gymnotid fish: Apteronotus leptorhynchus, Eigenmannia virescens and Sternopygus sp. Immunohistochemical data have revealed that substance P is never present in the afferent fibers but is specifically localized in the electroreceptors of the three species examined. Substane P immunoreactivity is strictly localized in the sensory cells of the ampullary organs of all three species and in those of the tuberous organs of Apteronotus leptorhynchus and Sternopygus sp. In contrast, weak substance P immunoreactivity was observed only in certain tuberous sensory cells of Eigenmannia. Substance P immunoreactivity was also found in the accessory cells of certain organs: it was detected in the two types of accessory cells of the tuberous organs of Eigemmannia virescens, in the accessory cells type 2 of the tuberous organs of Sternopygus sp., and in all accessory cells of ampullary organs of Sternopygus sp. and Apteronotus leptorhynchus. In Sternopygus sp., positive staining was only evident if the substance P antibody was used at low concentration. Immunoreactivity for substance P in the sensory cells suggests that it has a transmitter or modulator function in these electroreceptors; the presence of substance P in the accessory cells remains to be explained.     

Directional characteristics of tuberous electroreceptors in the weakly electric fish,Hypopomus (Gymnotiformes)

This paper is an electrophysiological study of the directionality of the tuberous electroreceptors of weakly electric fish. We recorded from two classes of tuberous electroreceptors known for pulse gymnotiforms: Burst Duration Coders (BDCs), and Pulse Markers (PMs). Both code for stimulus amplitude, although the dynamic range for BDCs is greater, and both exhibit strong directional preferences. Polar plots of spike number (for BDCs) or spike threshold (for PMs) versus electric field azimuth, are figure-8 shaped with two asymmetrical, elliptical lobes separated by 180°. The best azimuth of these two types of receptors from a given body region correlate with each other and with measures of best azimuth for transepidermal current flow. The shape and asymmetry of the directionality profiles appear to be caused by filter dynamics of the receptors. Pulse Markers are located on the anterior part of the body surface while Burst Duration Coders are located all over. The best directions of receptors in the anterior third of the body vary systematically with location from 0° to 180°. This region is probably critical for determining the direction of local electric fields. Together these receptors provide the CNS with sufficient information to construct a map of horizontal plane electric field directions.Abbreviations BDC Burst Duration Coder - ELL electrosensory lateral line lobe - EOD electric organ discharge - nALL anterior lateral line nerve - PM Pulse Marker     

Functional features of electroreceptors (small pit organs) in the catfish

Characteristics of responses of the small pit organs of the catfishIctalurus nebulosus to the action of electrical stimuli of varied polarity, intensity, and duration were studied. Single fibers of the lateral nerve innervating these organs possessed regular spontaneous activity with a frequency of 35–45/sec or grouped activity, coinciding with the rhythm of the animal's swimming movements. Threshold current densities varied from 10⁻¹¹ to 10⁻¹⁰ A/mm². Electrical stimuli evoked a phasic-tonic response of the receptor. The latent period was 10–50 msec for on-responses and 10–200 msec for off-responses. In the presence of strong electric fields the receptor responded to a cathodal stimulus by excitation, whereas under ordinary experimental conditions an anodal stimulus is excitatory. The properties of small pit organs are compared with the characteristics of other electroreceptors.     

Autocrine expression and ontogenetic functions of the PACAP ligand/receptor system during sympathetic development

The superior cervical ganglion (SCG) is a well-characterized model of neural development, in which several regulatory signals have been identified. Vasoactive intestinal peptide (VIP) has been found to regulate diverse ontogenetic processes in sympathetics, though functional requirements for high peptide concentrations suggest that other ligands are involved. We now describe expression and functions of pituitary adenylate cyclase-activating polypeptide (PACAP) during SCG ontogeny, suggesting that the peptide plays critical roles in neurogenesis. PACAP and PACAP receptor (PAC(1)) mRNA's were detected at embryonic days 14.5 (E14.5) through E17.5 in vivo and virtually all precursors exhibited ligand and receptor, indicating that the system is expressed as neuroblasts proliferate. Exposure of cultured precursors to PACAP peptides, containing 27 or 38 residues, increased mitogenic activity 4-fold. Significantly, PACAP was 1000-fold more potent than VIP and a highly potent and selective antagonist entirely blocked effects of micromolar VIP, consistent with both peptides acting via PAC(1) receptors. Moreover, PACAP potently enhanced precursor survival more than 2-fold, suggesting that previously defined VIP effects were mediated via PAC(1) receptors and that PACAP is the more significant developmental signal. In addition to neurogenesis, PACAP promoted neuronal differentiation, increasing neurite outgrowth 4-fold and enhancing expression of neurotrophin receptors trkC and trkA. Since PACAP potently activated cAMP and PI pathways and increased intracellular Ca(2+), the peptide may interact with other developmental signals. PACAP stimulation of precursor mitosis, survival, and trk receptor expression suggests that the signaling system plays a critical autocrine role during sympathetic neurogenesis.     

Regenerative capability in the hindlimb of Xenopus laevis during ontogenetic development

The regenerative capacity of hindlimb of Xenopus laevis was investigated by amputating the limbs at four levels in various developmental stages including younger postmetamorphosed froglets. Amputations of limbs were performed at the base of limb in stages 50, 51, 52, 53, 54, 55, 58, and 60 (Nieuwkoop and Faber's table), at the middle of limb bud in stages 50, 51, 52 and 54, and at mid-thigh and mid-shank in stages 58 and 60, and the froglets in 2 and 3 cm in snout-vent length. In the present experiments the regenerative capacity of limbs was expressed by the rate of regeneration and morphogenesis. Tadpoles in the stages after 55 failed to regenerate when the limbs were amputated at base level, but individuals in all the other experimental series exhibited regeneration in various rates irrespective of the level of amputation and the stage. The regenerative capacity increased distally along the proximo-distal axis of the limb when amputated at the same stage, while regeneration was better in younger stages than that in older stages when amputations were made at the same levels. The regenerates obtained by amputation of limbs in stages between 50 and 54, were mainly digitated in that they had 5 toes with 3 claws which is the same pattern with the normal limb, 4 toes with 2 claws, 3 toes with 2 claws or one, and 2 toes with one claw etc. Tadpoles at stage 50 could regenerate toes and claws without defect, but in the later the regenerative capacity gradually declined by reducing the number of toes and claws and accompanied by malformation of skeleton as the stage proceeded. The tadpoles in stages after 58, and the froglets of 2 and 3 cm, produced various types of heteromorphic regenerates of shapes such as cone, spike or rod of which the centra were occupied with cartilage rods. However these regenerates showed no morphological differences according to the developmental stages. These heteromorphic regenerates continued their growth even after one year without any sign of development of digitated feet.     

Structure-function relationships in the integument of Salamandra salamandra during ontogenetic development

Morphological, cytological and transport properties of the integument of Salamandra salamandra were investigated during natural ontogenetic development, from birth to adult. Three stages were operationally defined: I, larvae, from birth to metamorphosis; II, metamorphosis (judged externally by the colour change and loss of the gills); and III, post-metamorphosis to adult. Pieces of skin were fixed at various stages for immunocytochemical examinations, and the electrical properties were investigated on parallel pieces. Distinct cellular changes take place in the skin during metamorphosis, and lectin (PNA, WGA and ConA) binding indicates profound changes in glycoprotein composition of cell membranes, following metamorphosis. Band 3 and carbonic anhydrase I (CA I) were confined to mitochondria-rich (MR)-like cells, and were detected only in the larval stage. CA II on the other hand, was detected both in MR-like and in MR cells following metamorphosis. The electrical studies show that the skin becomes more tight (transepithelial resistance increases) upon metamorphosis, followed by manifestation of amiloride-sensitive short-circuit current (I(SC)) indicating that functional Na+ uptake has been acquired. The skin of metamorphosed adults had no finite transepithelial Cl- conductance, and band 3 was not detected in its MR cells. The functional properties of MR-like and MR cells remain to be established.     

Intermediate metabolism during the ontogenetic development of Anastrepha fraterculus (Diptera: Tephritidae)

The fruit fly Anastrepha fraterculus is a major pest of native and exotic fruit trees in South America. Changes in weight, water content and metabolism were observed during its ontogenetic development in standard conditions (25 degrees C, RH=60% and 14 h:10 h photoperiod). The metabolic variables glycogen, total proteins, triglycerides and total lipids were measured by means of spectrophotometric methods. The results were correlated with pupae metamorphosis, temporal pattern, and beginning of adult life. Pupae were observed daily, and a sub-sample of 10 individuals was collected and maintained at -20 degrees C. The same procedure was performed with adults at 4 days after adult eclosion. Levels of total lipids and triglycerides were constant during pupal development, peaking in 312-h-old pupae. In 0-h-old pupae, glycogen levels were high, and decreased progressively until the insects were 312 h old. The peak in total proteins coincides with the post-histolysis period of the larval tissue (96-120 h). These results indicated that glycogen and proteins may be the principal sources of energy for metamorphosis. Total lipid and triglyceride contents remained steady during metamorphosis, and these were consumed in the first 4 days of adult life.     

Prenatal and perinatal factors influencing nociception, addiction and behavior during ontogenetic development

This review, which summarizes our findings concerning the long-term effects of pre-, peri- and postnatal factors affecting development, nociception and sensorimotor functions, focuses on three areas: 1) perinatal factors influencing nociception in adult rats were examined in rats with hippocampal lesions, after the administration of stress influencing and psychostimulant drugs (dexamethasone, indomethacine and methamphetamine); 2) the effect of pre- and early postnatal methamphetamine administration was shown to impair the development of sensorimotor functions tested in rat pups throughout the preweaning period; 3) the effect of extensive dorsal rhizotomy of the brachial plexus during the early postnatal period was studied with respect to neuropathic pain development and sensorimotor functions. The present study indicates that prenatal or neonatal stress, as well as various drugs, may disturb the development of the nociceptive system and cause long-term behavioral changes persisting to adulthood and that some types of neuropathic pain cannot be induced during the first two postnatal weeks at all. A mature nervous system is required for the development of the described pathological behaviors.     

Differential expression of p63 isoforms in female reproductive organs

p63 is the identity switch for uterine/vaginal epithelial cell fate, and disruption of p63 expression by diethylstilbestrol (DES) induces cervical/vaginal adenosis in mice. In this article, we report the expression patterns of p63 isoforms (TA, DeltaN, alpha, beta and gamma) in mice, focusing on the reproductive tract. We also present the reproductive tract phenotype of female p63-/- mice. Finally, to better evaluate the potential role of p63 in human development of DES-induced cervical/vaginal adenosis, we describe the ontogeny of p63 in human female fetuses. In adult mice, the DeltaN isoforms of p63 were expressed only in squamous/basal/myoepithelial cells of epithelial tissues, while TA isoforms of p63 were highly expressed in germ cells of the ovary and testis. In fetal mice, the DeltaN and alpha forms of p63 were expressed in the cloacal and urogenital sinus epithelia. In the female p63-/- mice, the sinus vagina developed, but p63-/- sinus vaginal epithelium failed to undergo squamous differentiation confirming an essential role of p63 in squamous epithelial differentiation. Although TAp63 was highly expressed in developing primordial germ cells/oocytes, p63-/- ovaries and oocytes developed normally. The ontogeny of p63 in female reproductive organs was essentially identical in mouse and human. In the human fetus at the susceptible stage for DES-induced cervical/vaginal adenosis, most cervical/vaginal epithelial cells were columnar and negative for p63. Therefore, inhibition of p63 expression by DES should change the cell fate of human Müllerian duct epithelial cells and cause cervical/vaginal adenosis as previously demonstrated in mouse.     

Phenotypic plasticity, heterochrony and ontogenetic repatterning during juvenile development of divergent Arctic charr (Salvelinus alpinus)

Phenotypic plasticity is a developmental process that plays a role as a source of variation for evolution. Models of adaptive divergence make the prediction that increasing ecological specialization should be associated with lower levels of plasticity. We tested for differences in the magnitude, rate and trajectory of morphological plasticity in two lake populations of Arctic charr (Salvelinus alpinus) that exhibited variation in the degree of resource polymorphism. We reared offspring on diet treatments that mimicked benthic and pelagic prey. Offspring from the more divergent population had lower levels of morphological plasticity. Allometry influenced the rate of shape change over ontogeny, with differences in rate among ecomorphs being minimal when allometric variation was removed. However, plasticity in the spatial trajectory of development was extensive across ecomorphs, both with and without the inclusion of allometric variation, suggesting that different aspects of shape development can evolve independently.     

Time-dependent expression of cytochrome p450 epoxygenases during human prenatal development

There is growing evidence that some members of cytochrome P450 enzymes contribute to regulation of normal prenatal development. CYP epoxygenases (CYP2C and CYP2J subfamilies) convert arachidonic acid into four regioisomeric epoxyeicosatrienoic acids (EETs), biologically active molecules involved in mitogenesis and cell signaling. Almost nothing is known about localization of their expression in tissues during human prenatal development. The spatio-temporal expression pattern of CYP2C8, CYP2C9, CYP2C19 and CYP2J2 in human embryonic/fetal intestines, liver, and kidney was investigated by immunohistochemical method. CYP epoxygenases are expressed already in early stages of development in these embryonic/fetal tissues (as early as 7th week of IUD in the intestines, 5th week of IUD in the liver, and 6th week of IUD in the kidney). In kidney, CYP epoxygenases are expressed in the metanephrogenic blastema (but not in the uninduced mesenchyme) and in the tubular system. In the intestines, diverse CYP epoxygenases distribution along crypt-villus axis could suggest role in cell differentiation. Moreover, we detected higher CYP2J2 level in these organs than in adult tissue samples.     

Time-dependent expression of cytochrome p450 epoxygenases during human prenatal development

There is growing evidence that some members of cytochrome P450 enzymes contribute to regulation of normal prenatal development. CYP epoxygenases (CYP2C and CYP2J subfamilies) convert arachidonic acid into four regioisomeric epoxyeicosatrienoic acids (EETs), biologically active molecules involved in mitogenesis and cell signaling. Almost nothing is known about localization of their expression in tissues during human prenatal development. The spatio-temporal expression pattern of CYP2C8, CYP2C9, CYP2C19 and CYP2J2 in human embryonic/fetal intestines, liver, and kidney was investigated by immunohistochemical method. CYP epoxygenases are expressed already in early stages of development in these embryonic/fetal tissues (as early as 7th week of IUD in the intestines, 5th week of IUD in the liver, and 6th week of IUD in the kidney). In kidney, CYP epoxygenases are expressed in the metanephrogenic blastema (but not in the uninduced mesenchyme) and in the tubular system. In the intestines, diverse CYP epoxygenases distribution along crypt-villus axis could suggest role in cell differentiation. Moreover, we detected higher CYP2J2 level in these organs than in adult tissue samples.     

p107 and p130 Coordinately regulate proliferation,Cbfa1 expression,and hypertrophic differentiation during endochondral bone development

During endochondral bone development, both the chondrogenic differentiation of mesenchyme and the hypertrophic differentiation of chondrocytes coincide with the proliferative arrest of the differentiating cells. However, the mechanisms by which differentiation is coordinated with cell cycle withdrawal, and the importance of this coordination for skeletal development, have not been defined. Through analysis of mice lacking the pRB-related p107 and p130 proteins, we found that p107 was required in prechondrogenic condensations for cell cycle withdrawal and for quantitatively normal alpha1(II) collagen expression. Remarkably, the p107-dependent proliferative arrest of mesenchymal cells was not needed for qualitative changes that are associated with chondrogenic differentiation, including production of Alcian blue-staining matrix and expression of the collagen IIB isoform. In chondrocytes, both p107 and p130 contributed to cell cycle exit, and p107 and p130 loss was accompanied by deregulated proliferation, reduced expression of Cbfa1, and reduced expression of Cbfa1-dependent genes that are associated with hypertrophic differentiation. Moreover, Cbfa1 was detected, and hypertrophic differentiation occurred, only in chondrocytes that had undergone or were undergoing a proliferative arrest. The results suggest that Cbfa1 links a p107- and p130-mediated cell cycle arrest to chondrocyte terminal differentiation.     

Differential DNA methylation and gene expression during development of reproductive and vegetative organs in Ilex species

Journal of Plant Research - Differential epigenetic (DNA cytosine methylation) and gene expression patterns were investigated in reproductive and vegetative organs from Ilex paraguariensis and I....     

p38在小鼠着床前胚胎中的表达（简报）

探讨p38 MAPK在小鼠着床前胚胎期的表达图式,并对其作用作初步分析。用免疫印迹法分析胚胎全裂解物中的p38蛋白。为考察p38在着床前发育中的作用,在胚胎培养液中添加p38专一性抑制剂SB203580。此外对同位素标记的胚胎作双向电泳分析,示踪ZGA(zygotic gene activation,合子型基因激活)标志物TRC的表达情况。在卵母细胞中能检测到低水平的p38蛋白,而在合子中的检测度更低,表明p38是贮存于卵母细胞内的母型转录物,自减数分裂期随其它母型转录物一起逐步降解。到2细胞中期p38蛋白的表达量开始恢复,在4细胞时达到顶峰,在8细胞时又跌落。D38蛋白在2到4细胞期的表达量上升提示该蛋白在小鼠着床前胚胎发育中可能发挥一定作用。经与p38抑制剂SB203580共培养后的2细胞中期胚胎中仍能清晰检测到TRC,因而以TRC为标志的ZGA对SB203580不敏感。SB203580同样不能阻止胚胎发育到桑椹胚期。