A strategy for genetic improvement of Barbados Blackbelly sheep in the Caribbean in the context of small flock sizes.

This paper points out that several breeds of prolific sheep in the world possess a major gene for prolificacy. However, the existence of such a major gene in the prolific breeds of hair sheep in the Caribbean remains to be researched. This paper assumes that this gene (F) exists in the Barbados Blackbelly sheep and goes on to outline a strategy to exploit it for the benefit of the small farmer as well as the large private producer. The strategy aims at developing one type of sheep for the small farmer by selective reduction in the frequency of the F gene thereby reducing the frequency of multiple births. The development of another type of highly prolific sheep is suggested for use by the large producer through selective increase in the frequency of the F gene. Large producer is expected to employ high levels of feeding and management including artificial rearing of lambs so as to minimise mortality of multiplets. The paper further discusses within breed selection for genetic improvement in general fitness of hair sheep. In this connection, the development of an open nucleus flock is suggested involving cooperation among participant smallholders.

A strategy for genetic improvement of Barbados Blackbelly sheep in the Caribbean in the context of small flock sizes.

This paper points out that several breeds of prolific sheep in the world possess a major gene for prolificacy. However, the existence of such a major gene in the prolific breeds of hair sheep in the Caribbean remains to be researched. This paper assumes that this gene (F) exists in the Barbados Blackbelly sheep and goes on to outline a strategy to exploit it for the benefit of the small farmer as well as the large private producer. The strategy aims at developing one type of sheep for the small farmer by selective reduction in the frequency of the F gene thereby reducing the frequency of multiple births. The development of another type of highly prolific sheep is suggested for use by the large producer through selective increase in the frequency of the F gene. Large producer is expected to employ high levels of feeding and management including artificial rearing of lambs so as to minimise mortality of multiplets. The paper further discusses within breed selection for genetic improvement in general fitness of hair sheep. In this connection, the development of an open nucleus flock is suggested involving cooperation among participant smallholders.

Production performance of blackhead persian sheep in an experimental flock in Tobago, West Indies

The effect of lambing season-year, sex and type of birth on lamb weights at birth, weaning at 56 days and 6-months, and daily gain and preweaning mortality was studied. Data were based on 131 lambs from 103 lambings by Blackhead Persian ewes in 1982-85 at the Blenheim Sheep Station in Tobago (West Indies). The effect of the lambing season-year on measures of ewe reporductive performance (number of lambs borm total, born alive and, weaned per ewe lambing) and total lamb weight weaned was also studied. The average values for lamb body weights at birth, weaning and 6-month, and daily gain mortality were: 2.57 kg, 10.9kg, 16.6kg, 147g and 14.6 percent respectively. The flock means for number of lambs born total, born alive, weaned and for total weight of lambs weaned at 56 days were 1.27, 1.24, 1.08 and 13.2 kg, respectively. The lambing season-year effects caused significant variation in body weights at birth and 6-month, and lamb mortallity only. Sex of lamb had no significant effect on any of the traits studied. The effect of type of birth was apparent for all traits except 6-month body weight and lamb mortality. Single born lambs. Ewes with twins weaned 54 percent more lamb than ewes with singles (16.0 vs. 10.4 kg). Lambs with below average birth weight had significantly higher mortality than those with above average birth weight (20.7 vs 8.5 percent)(AU)

Production performance of Barbados blackbelly sheep in Tobago, West Indies.

Data on 298 lambs from 161 ewes of the Barbados Blackbelly breed from 1982 to 1985 at the Blenheim Sheep Station in Tobago were studied to determine the influence of lambing season-year, sex and type of birth on body weights of lambs at birth, at 56 days (weaning) and 6 months of age, preweaning daily gain and mortality. The effect of the lambing season-year on litter size (total number of lambs born, born alive and weaned per ewe lambing) and total lamb weight at weaning were examined. The flock averages for body weights of lambs at birth, at 56 days and 6 months of age, preweaning daily gain and mortality were 2.75, 10.8, 19.2, 0.152kg, and 18.3%, respectively. Averages for total number of lambs born, born alive and weaned were 1.92, 1.88, 1.51, respectively, and total lamb weight at weaning was 16.7kg. The lambing season-year effect was apparent for all traits studied except for litter size and lamb mortality. Sex of lamb was not a significant source of variation. Type of birth had significant effect on lamb growth traits and total lamb weight at weaning. Single born lambs were heavier than those born as twins or more. Ewes with triplets or more weaned 21kg lamb weight compared to 18 and 12kg for twins and singles, respectively. Lamb mortality was 27.8% for those with below average birth weight and 8.8% for those with above average birth weight (P<0.05).

FMRFamide in the amphibian brain: a comprehensive survey.

Mapping of FMRFamidergic neural circuitry in the amphibian brain has been done by immunohistochemical methods. Comparative evidence suggests that there are similarities and differences in the overall pattern of distribution of FMRFamide-ir elements in the brain among the three amphibian orders and within each order. FMRFamide is expressed in neurons in some circumscribed areas of the brain. A part of these neurons is concentrated in classical neurosecretory areas of the hypothalamus in a bilaterally symmetrical fashion. Similar neurons occur occasionally in the midbrain, but are virtually absent from the hindbrain. Anurans are unique among amphibians to show FMRFamide neurons in the medial septum and diagonal band of Broca. A viviparous gymnophione is known to possess a small population of such neurons in the dorsal thalamus. Together, the FMRFamide neurons contribute to an extensive fiber network throughout the amphibian brain. Descriptive developmental studies suggest that the rostral forebrain-located FMRFamide neurons originate in the olfactory placode and then migrate into the brain along the route of the vomeronasal-olfactory-terminal nerve complex. Olfactory placodal ablation in an anuran and a urodele provide experimental support to this contention. Other FMRFamide neuronal cell groups, in the hypothalamus and dorsal thalamus, are supposed to arise from non-placodal precursors. The neuroanatomical distribution (projection of immunoreactive processes to areas of the fore-, mid-, and hindbrain as well as to cerebrospinal fluid, co-localization with other neuropeptides, and presence in the median eminence) has furnished morphological correlates of possible functions of FMRFamide in the amphibian CNS. While amphibian FMRFamide-like or structurally related peptides remain to be isolated and characterized, the sum of the distribution pattern of FMRFamide-like immunoreactivity suggests that it may act as a neurotransmitter or a neuromodulator, and also may have endocrine regulatory functions.

Localization of FMRFamide-like immunoreactivity in the brain of the viviparous skink (Chalcides chalcides).

Neuroanatomical distribution of FMRFamide-like immunoreactivity was investigated in the brain and olfactory system of the viviparous skink, Chalcides chalcides. In the adult brain FMRFamide immunoreactive (ir) perikarya were observed in the diagonal band of Broca, medial septal nucleus, accumbens nucleus, bed nucleus of the anterior commissure, periventricular hypothalamic nucleus, lateral forebrain bundle, and lateral preoptic, subcommissural, suprachiasmatic and lateral hypothalamic areas. This pattern was seen in both male and female brains. Though all major brain areas showed FMRFamide-ir innervation, the densest ir fiber network was observed in the hypothalamus. During development, ir elements were observed for the first time in embryos at mid-pregnancy. FMRFamide perikarya were located along the ventral surface of the vomeronasal nerve, in the olfactory peduncle mediobasally, as well as in the anterior olfactory nucleus and olfactory tubercle. Furthermore, some ir neurons were observed in the rhombencephalic reticular substance; however, the ir fiber network was poorly developed. Later in development FMRFamide-ir neurons appeared also in the bed nucleus of the anterior commissure as well as the rhombencephalic nucleus of solitary tract and the dorsal motor nucleus of vagus nerve. In juveniles, the distribution profile of FMRFamide immunoreactivity was substantially similar to that of the adults, with a less widespread neuronal distribution and a more developed fiber network. Ontogenetic presence of FMRFamide immunoreactivity in the nasal area has been linked to the presence of a nervus terminalis in this reptile.

Development and distribution of FMRFamide-like immunoreactivity in the toad (Bufo bufo) brain.

By using immunohistochemistry, we studied the development and distribution of the FMRFamide-like immunoreactive (ir) neuronal system in the toad brain during the ontogeny. In addition to this, experimental evidence was provided to show that the rostral forebrain-located FMRFamide neurons originate in the olfactory placode and then migrate into the brain along the olfactory pathway. During early development, within the brain, FMRFamide-ir perikarya first appeared in the periventricular hypothalamus. Later in development, FMRFamide-ir cells were visualized in the rostralmost forebrain simultaneously with similar ir cells in the developing olfactory mucosa. Selective ablation of the olfactory placode(s), prior to the appearance of the first FMRFamide-ir cells in the brain, resulted in the total absence of ir cells in the telencephalon (medial septum and mediobasal telencephalon) of the operated sides(s). The preoptic-suprachiasmatic-infundibular hypothalamus-located FMRFamide-ir neurons were not affected by olfactory placodectomy, arguing that they do not originate in the placode. This result points to the placode as the sole source of such neurons in the rostral forebrain.

Ontogeny of pituitary adenylate cyclase-activating polypeptide (PACAP) in the frog (Rana ridibunda) tadpole brain: immunohistochemical localization and biochemical characterization.

The anatomic distribution and biochemical characteristics of the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) were investigated in the central nervous system of the frog, Rana ridibunda, during development. Three to four days after hatching, at stages IV-VII, PACAP-immunoreactive perikarya were detected in the dorsal thalamus within the anterior ventral area, and a few fibers were found in the medial pallium. Positive cell bodies were first observed in the hypothalamus at stages VIII-IX, at the level of the dorsal and ventral infundibular nuclei. In these regions, the number of positive perikarya increased during ontogeny. In tadpoles, during the mid- and late premetamorphosis, a more complex organization of the PACAP-immunoreactive system was found in the thalamus with the appearance, at stages IX-XII, of two additional groups of positive neurons in the ventrolateral area and posterocentral nucleus. At stages XIII-XVIII of larval development and subsequent larval stages, PACAP-immunoreactive fibers were found in the median eminence. In newly metamorphosed animals, several additional groups of positive perikarya appeared in the medial pallium, the preoptic nucleus, the torus semicircularis, the tegmentum of the mesencephalon, and the cerebellum. The immunoreactive peptide contained in the tadpole brain was characterized by high performance liquid chromatography analysis combined with radioimmunoassay quantification. At all stages investigated, the predominant form of PACAP-immunoreactive material coeluted with synthetic frog PACAP38. The occurrence of PACAP soon after hatching indicates that the peptide may exert neurotrophic activities. The existence of immunoreactive elements in several thalamic regions at mid- and late premetamorphic stages suggests that PACAP may act as a neurotransmitter, neuromodulator, or both, during ontogenesis. Finally, the presence of PACAP-immunoreactive perikarya in hypothalamic nuclei and nerve fibers in the median eminence supports the view that PACAP may play a role in the control of pituitary hormone secretion during larval development.

Comparative immunocytochemical study of FMRFamide neuronal system in the brain of Danio rerio and Acipenser ruthenus during development.

The distribution of FMRFamide-like immunoreactive (ir) neurons and fibers was investigated in the central nervous system of developing zebrafish and juvenile sturgeon (sterlet). Adult zebrafish was also studied. In zebrafish embryos FMRFamide-ir elements first appeared 30 h post-fertilization (PF). Ir somata were located in the olfactory placode and in the ventral diencephalon. FMRFamide-ir fibers originating from diencephalic neurons were found in the ventral telencephalon and in ventral portions of the brainstem. At 48 h PF, the ir perikarya in the olfactory placode displayed increased immunoreactivity and stained fibers emerged from the somata. At 60 h PF, bilaterally, clusters of FMRFamide-ir neurons were found along the rostro-caudal axis of the brain, from the olfactory placode to rostral regions of the ventro-lateral telencephalon. At 60 h PF, numerous ir fibers appeared in the dorsal telencephalon, optic lobes, optic nerves, and retina. Except for ir fibers in the hypophysis at the age of 72 h PF, and a few ir cells in the nucleus olfacto-retinalis (NOR) at the age of 2 months PF, no major re-organization was noted in subsequent ontogenetic stages. The number of stained NOR neurons increased markedly in sexually mature zebrafish. In adult zebrafish, other ir neurons were located in the dorsal zones of the periventricular hypothalamus and in components of the nervus terminalis. We are inclined to believe that neurons expressing FMRFamide originate in the olfactory placode and in the ventricular ependyma in the hypothalamus. On the same grounds, a dual origin of FMRFamide-ir neurons is inferred in the sturgeon, an ancestral bony fish: prior to the observation of ir cells in the nasal area and in the telencephalon stained neurons were noted in circumventricular hypothalamic regions.

Mammalian and chicken I forms of gonadotropin-releasing hormone in the gonads of a protochordate, Ciona intestinalis.

Two forms of gonadotropin-releasing hormone (GnRH) were isolated from the gonads of the tunicate, Ciona intestinalis. The primary structure of the purified peptides was determined by MS and chemical sequence analysis. Both GnRH forms have blocked NH(2) and COOH termini, and their primary structures are identical to mammalian (mGnRH) and chicken I (cGnRH-I) forms reported previously in vertebrates. A total of 1.2 mg of purified cGnRH-I and 0.98 mg of mGnRH was obtained from 100 g of Ciona gonads. The physiological effects of native GnRHs included the induction of synthesis and secretion of sex steroids from ciona gonads and the secretion of luteinizing hormone from rat pituitary. These results suggest that the primary structure and functional roles of mGnRH and cGnRH-I have been highly conserved throughout evolution of chordates.

Ontogenetic profile of FSH and LH in Rana esculenta.

Circulating levels and pituitary content of FSH and LH were determined by specific radioimmunoassays in Rana esculenta starting a few days after hatching until the completion of metamorphosis. Both gonadotropins were found in the pituitary as well as in the blood plasma at all stages of development examined here. The plasma concentrations of FSH and LH were more or less uniform during pre- and prometamorphosis, but increased significantly at the onset of metamorphic climax. The plasma levels of FSH and LH remained high at the completion of metamorphosis. The pituitary content of FSH and LH was low in early premetamorphosis. It increased slightly through prometamorphosis and metamorphic climax, following which a highly significant increase occurred. Whereas plasma concentrations of FSH and LH were essentially similar within a single stage of development, the pituitary FSH content was severalfold higher than pituitary LH. The significance of these results is discussed in relation to the functional maturation of the brain-pituitary-gonadal axis in the frog.

Distribution of FMRFamide-like immunoreactivity in the amphibian brain: comparative analysis.

FMRFamide is a small neuropeptide present in particular neurons of the basal forebrain and midbrain of the vertebrate groups studied, especially fishes and mammals. In order to assess interspecies variation, the distribution of FMRFamide-like immunoreactivity was studied in the brains of 13 species of amphibian. Although FMRFamide-immunoreactive (IR) terminals occurred throughout much of the brain, IR cell groups were noted in circumscribed regions of the CNS. In the eight anuran species studied, two major populations of labeled perikarya were observed: one in the septopreoptic area and another one in the caudal portion of the diencephalon. The rostrocaudal extent of both and the number of labeled somata in each neuronal group displayed species-specific differences. In urodeles and gymnophiones, labeled perikarya were located in the diencephalon, but there were remarkable species differences in the number of such cells. It is discussed whether sex or season of collection may account for some of the differences observed. The distribution of FMRFamide-IR perikarya, fibers, and pathways in the brain of anurans, urodeles, and gymnophiones was compared. The existence of FMRFamide perikarya in the anterior preoptic neuropil and medial septum appeared to be a feature common to all anurans; labeled neurons in the dorsal thalamus, however, may be present only in the (viviparous) gymnophione Typhlonectes compressicauda. Cerebrospinal fluid contacting FMRFamide neuronal cell bodies and fibers were observed in each of the three taxonomic orders. The data are compared with those previously obtained for other groups of vertebrates.

Distribution of vasoactive intestinal peptide-like immunoreactivity in the brain and pituitary of the frog (Rana esculenta) during development.

The localization of vasoactive intestinal peptide (VIP)-like immunoreactive (ir) elements was investigated in the brain of the anuran amphibian, Rana esculenta, during development. Using an antiserum raised against the porcine VIP, ir cell bodies and fibers were observed in the forebrain of tadpoles a few days after hatching. During early premetamorphosis, ir perikarya were distributed in the ventral infundibular nucleus of the hypothalamus and in the posterocentral nucleus of the thalamus. Labeled fibers were detected in the olfactory bulbs and in the hypothalamus. In these larvae, furthermore, several VIP-ir cells were found in the pars distalis of the pituitary and there were ir fibers in the pars nervosa. In tadpoles at stages VIII-IX, a new group of VIP-labeled neurons was observed in the dorsal part of the infundibular nucleus. In other brain regions, the distribution of the immunoreactivity was similar to that described in the earliest stages, i.e., IV-VII. During mid-premetamorphosis, stages X-XII of development, an additional set of ir perikarya appeared in the ventrolateral area of the thalamus. During late premetamorphosis, stages XIII-XVIII, the organization of VIP-like immunoreactivity was more complex and its distribution more widespread. Two new groups of ir cell bodies appeared, one in the preoptic nucleus and another in the anteroventral area of the thalamus, and for the first time, VIP immunoreactivity was observed in the median eminence. This distribution pattern persisted through to the prometamorphic, four-limb stage. Strikingly, no VIP-ir elements were observed anywhere in the mid- and hindbrain. The present results indicate that a VIP-like ir peptide may be involved in the processing of olfactory information or may act as a neurohormone, hypophysiotropic factor, and neuromodulator in the brain of R. esculenta during development.

Comparative analysis of FMRFamide-like immunoreactivity in caiman (Caiman crocodilus) and turtle (Trachemys scripta elegans) brains.

The distribution of FMRFamide (FMRFa)-like peptides in caiman (Caiman crocodilus) and turtle (Trachemys scripta elegans) brains was studied by immunohistochemistry. In both species, distinct groups of FMRFa-like immunoreactive (ir) perikarya were present in the medial septal nucleus, accumbens nucleus, nucleus of the diagonal band of Broca, suprachiasmatic area, lateral hypothalamic area, and periventricular hypothalamic nucleus. A few FMRFa-ir neurons in the hypothalamic area were located in the neuroepithelial cell lining of the third ventricle. FMRFa-ir fibers were scattered in all major areas of the brain, from the olfactory bulbs to the rhombencephalon. They formed dense aggregates in the medial septal area, basal telencephalon, median eminence, and infundibulum, and adjacent to the fourth ventricle. The most obvious difference between the FMRFa-ir systems in caimans and turtles concerned the number of nuclei that contained neurons with this immunoreactivity. Eight such clusters were present in the caiman brain, whereas thirteen clusters were found in the turtle brain. The turtle also displayed scattered FMRFa-ir somata in the anterior olfactory nucleus, striatum, lateral septal nucleus, medial and lateral cortex, medial forebrain bundle, lateral preoptic area, and lateral geniculate nucleus. In the caiman brain, a few FMRFa-ir neurons were noted in the ventrolateral area of the pallial commissure and an even smaller number of ir neurons was found dispersed in the optic tracts. Neither formed nuclear aggregates. The results are compared with those described for other vertebrates.

Comparative analysis of GnRH neuronal systems in the amphibian brain.

We have investigated the GnRH-ir neuronal systems in the brain of the oviparous urodele, Triturus vulgaris, ovoviviparous urodele, Salamandra salamandra, and viviparous caecilian, Typhlonectes compressicauda, and have reexamined Xenopus laevis, Ambystoma mexicanum, and Rana esculenta. Results showed that mGnRH neuronal system was diffused along the medioventral telencephalon and diencephalon with the numerical preponderance of GnRH cell bodies in the rostral mediobasal telencephalon in T. vulgaris and S. salamandra and in medial septal area and preoptic area respectively in Typhlonectes compressicauda and X. laevis. The cGnRH-II-ir perikarya were restricted to the midbrain tegmentum in X. laevis and T. compressicauda. In T. vulgaris, two distinct groups of cGnRH-II neurons were distinguished, one in the midbrain tegmentum and another in the paraventricular organ. The former was composed of comparatively bigger perikarya than the latter. In X. laevis brain, besides those in the rostralmost dorsomedial and ventromedial telencephalon and septopreoptic area, mGnRH neurons were also found in the habenulae and habenular commissure as well the infundibular hypothalamus. In A. mexicanum, reexamined, the preoptic area-located mGnRH neurons were distributed in the ependymal lining of the preoptic recess. In this neotenic urodele, furthermore, cGnRH-II neurons were also present in the rhombencephalon, as well as in the infundibular hypothalamus. It is thus clear that while GnRH-ir cell bodies are distributed in the fore-, mid- and hindbrain, their precise neuroanatomical localization varies somewhat within and among groups. Altogether, it is evident that mGnRH neuronal system is confined mainly to the forebrain, whereas cGnRH-II system is commonly found in the mid- and hindbrain. Additional morphological investigations are required to eventually define the functional neuroanatomy of GnRH in the amphibian brain.

Organization of atrial natriuretic factor-like immunoreactive system in the brain of the frog Rana esculenta during development.

Immunocytochemical distribution of the atrial natriuretic factor (ANF) has been studied in the brain and pituitary of the anuran Rana esculenta during development and in juvenile animals. Using human ANF and rat alpha-ANF antisera, immunoreactive cell bodies and nerve fibers were revealed in stage II-III tadpoles and in successive larval stages. Soon after hatching, stages II-III, the ANF-like-immunoreactive elements were confined to the preoptic area-median eminence complex. During successive stages of development, new groups of ANF-immunoreactive cell bodies appeared. In larval stage VI, immunoreactive perikarya were found in the rostral part of the anteroventral area of the thalamus and numerous ANF-like-immunoreactive cells appeared in the pars distalis of the pituitary. In larval stages XIV and XVIII, the distribution of ANF immunoreactivity was virtually similar. The ANF-immunoreactive cells in the preoptic nucleus and in the pituitary pars distalis were comparatively more abundant than in stage VI. During the metamorphic climax (stages XXI-XXII), a new group of ANF-immunoreactive cell bodies appeared in the rostral part of the ventrolateral area of the thalamus. During this stage, ANF-immunoreactive fiber projections were found in the pars intermedia for the first time. However, the pars distalis cells were very weakly immunofluorescent. The pattern of ANF immunoreactivity in the brain of juvenile animals was very similar to that described for stages XXI and XXII, whereas the pars distalis cells showed no immunoreactivity. It is conceivable that, early during development, ANF-related peptides may be involved in the regulation of pituitary secretion by means of autocrine mechanisms or may act as a classic pituitary hormone.

Distribution of somatostatin-like immunoreactivity in the brain of the frog, Rana esculenta, during development.

The anatomical distribution of somatostatin-like immunoreactivity in the central nervous system of the frog, Rana esculenta, during development and in juvenile specimens was investigated by indirect immunofluorescence. Soon after hatching, at stages II-III, somatostatin-like immunoreactive structures were found in the preoptic-median eminence complex. In stage VI tadpoles, new groups of immunopositive perikarya and nerve fibers appeared in the diencephalon, within the ventral infundibular nucleus and in the ventral area of the thalamus, as well as in the medial pallium. In stages XII-XIV of development, immunopositive perikarya were also present in the dorsal infundibular nucleus of the hypothalamus and ventrolateral area of the thalamus. A small group of somatostatin-like immunoreactive neurons appeared in the posteroventral nucleus of the rhombencephalon. However, these neurons were not seen in later stages of development. Tadpoles in stages XVIII, XXI-XXII and in juveniles were characterized by a wider distribution of immunoreactive cell bodies and fibers in the pallium. New groups of immunoreactive neurons were found in the dorsal and lateral pallium. The presence of positive perikarya in the lateral pallium is a transient expression found only in these stages. The organization of the somatostatinergic system was most complex during the metamorphic climax, with the appearance of positive cell bodies in the posterocentralis area of the thalamus, and in juvenile animals with the presence of perikarya in the ventral part of the medial pallium and within the central grey rhombencephali. In contrast to the adult frog, somatostatin neurons were not observed in the mesencephalon of tadpoles and juveniles.

Distribution of gonadotropin-releasing hormone immunoreactivity in the brain of Ichthyophis beddomei (Amphibia: Gymnophiona).

From a comparative viewpoint, we have investigated the presence and neuroanatomical distribution of gonadotropin-releasing hormone (GnRH)-immunoreactive material in the brain of a gymnophione amphibian, Ichthyophis beddomei. Immunocytochemical analysis of the adult brain and terminal nerves in both sexes shows the presence of neurons and fibers containing mammalian GnRH (mGnRH)- and chicken GnRH-II (cGnRH-II)-like peptides. With respect to GnRH-immunoreactive material, there are two distinct neuronal systems in the brain: one containing mGnRH, which is located in the forebrain and terminal nerve, and the other containing cGnRH-II, which is restricted to the midbrain tegmentum. Basically, this distribution pattern parallels that of many species of anurans and a urodele. Whereas the presence of cGnRH-II-immunoreactive fibers in the dorsal pallium of L. beddomei is a feature in common with a urodele amphibian, the total absence of cGnRH-II-like material in the median eminence is unique to this species. It is suggested here that the distribution profile of GnRH-like material within the brain and terminal nerve of I. beddomei represents a primitive pattern.

Sex and reproductive status related brain content of mammalian and chicken-II GnRHs in Rana esculenta.

Mammalian and chicken-II forms of gonadotropin-releasing hormone (mGnRH and cGnRH-II, respectively) have been measured simultaneously in the brain, pituitary, and peripheral terminal nerves (nasal area tissue) of adult males and females of a representative amphibian, Rana esculenta, during the annual reproductive cycle. Whereas in the male, brain concentrations of both GnRH forms showed significant reproductive status-related fluctuations, in the female brain only cGnRH-II content showed significant changes. The highest GnRH levels were recorded just prior to breeding in both sexes. In the pituitary both GnRHs were present in all seasons. In the peripheral terminal nerves, instead, only mGnRH was detected in all seasons confirming our previous immunohistochemical data. In both sexes furthermore, the brain and pituitary mGnRH levels were consistently much higher than those of cGnRH-II and there were no sex-related differences in the brain and pituitary content of GnRHs. Seasonal changes in brain GnRH levels may correlate with plasma sex steroid levels reinforcing the postulate that sex steroids affect GnRH neuronal systems.

Immunohistochemical demonstration of FSH and LH in the pituitary of the developing frog, Rana esculenta.

The ontogenetic pattern of immunohistochemically detectable FSH beta and LH beta cells was investigated in the pars distalis of the pituitary of the frog, Rana esculenta. The appearance, distribution, and percentage of these cells were examined in tadpoles from soon after hatching to the end of metamorphosis and in juveniles. We used monoclonal antibodies against bullfrog FSH beta and LH beta for single staining, and either mouse anti-bullfrog LH beta + guinea pig anti-rat FSH beta or rabbit polyclonal anti-bullfrog LH beta + mouse monoclonal anti-bullfrog FSH beta for double staining. The first appearance of gonadotropes, immunopositive for FSH beta, was revealed in stage 26 tadpoles. In successive stages of development the percentage of FSH beta-positive cells increased progressively and significantly. The mean percentage of these in the pars distalis cells increased from 0.7% in stage 26 to nearly 10% during the metamorphic climax (stages 31-33). In juveniles, the mean percentage of FSH beta-positive cells increased more than twofold compared to the climax value. The appearance of LH beta-positive cells was first recorded during the climax, and the mean percentage of LH beta-positive cells in juveniles reached levels as high as 30% or more, exceeding the number of FSH beta-positive cells. In climax, all LH beta-positive cells stained with anti-FSH beta as well. In juveniles, however, up to 80% of gonadotropes demonstrated colocalization of FSH beta and LH beta staining. We argue that both gonadotropins may be synthesized in all gonadotropes, and a small number of cells immunoreactive to either of the two gonadotropins may simply indicate that at that particular moment the cell contained detectable amounts of only one form of gonadotropin. These observations are discussed in relation to the possible involvement of hypothalamic influence in the differentiation of gonadotropes of the pituitary.