Organization of diencephalon of the sturgeons. External nuclei of the pretectal area

Cytoarchitectonics of the external nuclei of the pretectal area was studied in four species of the sturgeons: great sturgeon, Huso huso L., Kura sturgeon, Acipenser gueldenstaedtii persicus n. Kurensis Belyaeff, stellate sturgeon, Ac. stellatus Pall. and barbel sturgeon, Ac. nudiventris Lov. Study of morphological organization of the structures was carried out using routine Nissl staining and Bilschowski impregnation technique in Viktorov’s modification. Similar structure of this pretectal area was found in the species. Five nuclear formations were described in this pretectal area: parvocellular and magnocellular external pretectal nuclei, laminiform nucleus of pretectum, dorsal pretectal nucleus, and the accessory optic nucleus. Comparison of the obtained data with the literature revealed a high level of differentiation of the area of pretectum in the sturgeons. Also, a general conclusion is given on the organization of the whole pretectal area in the sturgeons.     

Periventricular and central nuclei of the pretectal area of diencephalon of the sturgeons

Cytoarchitectonics of periventricular and central nuclei of the pretectal area was studied in four species of the sturgeons: the great sturgeon Huso huso, L., the Russian sturgeon Acipenser gъldenst?dti persicus n. kurensis, Belyaeff, the starred sturgeon Ac. stellatus, Pall., and the barbel sturgeon Ac. nudiventris, Lov.; this pretectum part has a similar structure. Study of these parts of the pretectal area was carried out by methods of Nissl and Bielshowskii modified by Viktorov. In this part of the pretectal area, nine nuclear structures were described, eight of them—nuclear; these are ventral periventricular pretectal nucleus and its dorsal component, dorsal periventricular pretectal nucleus, nucleus of medial longitudinal bundle, subcomissural organ, medial and lateral intercalate nuclei, and central and posterior pretectal nuclei. The main attention has been paid to the issue of the evolutional progression of this part of the pretectal area in the sturgeons as compared with other Actinopterygii.     

Organization of hypothalamic area of diencephalon in the sturgeons

Cytoarchitectonics of hypothalamic area of diencephalon of the sturgeons was studied in serial sections by techniques of Nissl staining and Bielschowski impregnation in Viktorov’s modification. The hypothalamus was shown to be the most expanded area of diencephalon and forms its the most ventral part. The hypothalamic area of four studied sturgeons, the hausen, Huso huso L., Kura sturgeon, Acipenser guldenstaedtii persicus n. Kurensis Belyaeff, Caspian sturgeon, Ac. stellatus Pall. and the barbel sturgeon Ac. nudioventris Lov. was found to have similar structure. Eleven nerve structures are identified and described in the hypothalamic area: dorsal, ventral, and caudal periventricular zones, rostral and dorsal hypothalamic nuclei, ventral and ventrolateral hypothalamic nuclei, diffuse and central nuclei of the inferior lobes, nucleus of the vascular sac, and mammillary nucleus. Peculiarities and common features of organization of four major parts of hypothalamus of the sturgeons are considered in comparison with those of other ray-finned fish. The performed analysis indicates a high level of development of hypothalamus of the sturgeons.     

Outer nuclei of the diencephalon pretectal area of the sturgeons

Cytoarchitectonics of outer nuclei of the pretectal area was studied in four species of the sturgeons: the great sturgeon Huso huso, L., the Russian sturgeon Acipenser güldenst?dti persicus n. kurensis, Belyaeff, the starred sturgeon Ac. stellatus, Pall., and the barbel sturgeon Ac. nudiventris, Lov. Morphological organization of these structures was studied by methods of Nissl and Bielshowskii modified by Viktorov. In this part of the pretectal area, five nuclear structures were described: parvicellular and magnocellular outer pretectal nuclei, lamellar pretectal nucleus, and dorsal pretectal nucleus, as well as accessory optic nucleus. Comparison of the obtained results with literature data has revealed a sufficiently high level of differentiation of this pretectum part of the sturgeons. Besides, a general conclusion is presented on organization of their entire pretectal area.     

Periventircular and central nuclei of the diencephalon pretectal area of the sturgeons

Cytoarchitectonics of periventricular and central nuclei of the pretectal area was studied in four species of the sturgeons: the great sturgeon Huso huso, L., the Russian sturgeon Acipenser güldenst?dti persicus n. kurensis, Belyaeff, the starred sturgeon Ac. stellatus, Pall., and the barbel sturgeon Ac. nudiventris, Lov.; this pretectum part has a similar structure. Study of these parts of the pretectal area was carried out by methods of Nissl and Bielshowskii modified by Viktorov. In this part of the pretectal area, nine nuclear structures were described, eight of them--nuclear; these are ventral periventricular pretectal nucleus and its dorsal component, dorsal periventricular pretectal nucleus, nucleus of medial longitudinal bundle, subcomissural organ, medial and lateral intercalate nuclei, and central and posterior pretectal nuclei. The main attention has been paid to the issue of the evolutional progression of this part of the pretectal area in the sturgeons as compared with other Actinopterygii.     

Organization of diencephalon of the sturgeons. Posterior tubercular area. Caudal region

Cytoarchitectonics of caudal region of the posterior tubercular area of diencephalon was studied in four species of the true sturgeons: the giant sturgeon, Huso huso L., Kura sturgeon, Acipenser güldenstädtii persicus n. Kurensis Belyaeff, stellate sturgeon, Ac. stellatus Pall. and the barbel sturgeon Ac. nudiventris Lov. Morphologic organization of the brain region was studied by using staining techniques of Nissl and Bielschowski (Viktorov’s modification). Five nuclei were identified in the caudal region of the posterior tubercular area in all studied species: caudomedial, caudal and lateral thalamic nuclei forming the medial group, as well as lateral torus and posterior thalamic nuclei forming the lateral group. The latter two nuclei have their subdivisions. Characteristic peculiarity of the caudal region is the presence of only migrated nuclei. It is concluded that the level of differentiation of the diencephalon posterior tubercular area of the sturgeons allows considering it as one of the most evolutionary advanced structures among those in the actinopterygian fish line.     

Organization of the diencephalon ventrothalamic area in the sturgeons

Cytoarchitectonics of ventrothalamic area of the diencephalon of the sturgeons was studied in four species of the sturgeons, the great sturgeon Huso huso L., the Kura sturgeon Acipenser guelden-staedtii persicus n. Kurensis Belyaeff, the Caspian sturgeon, Ac. stellatus Pall., and the barbel sturgeon Ac. nudioventris Lov. using Nissl staining and Bilschowski impregnation techniques in Viktorov’s modification. The ventral thalamus of the sturgeons was found to include seven nuclei: ventromedial, ventrolateral, intermediate and ventral thalamic nuclei, thalamic eminence, and nuclei of ventral and medial optic tracts. The main attention was paid to comparative analysis of this area in the sturgeons and other actinopterygian fish. The structures common to all ray-finned fish or only to the lower teleosts and ganoids as well as characterizing only the sturgeons.     

Organization of diencephalon of the sturgeons. Posterior tubercular area. Periventricular and rostral parts

Cytoarchitectonics of periventricular and rostral parts of the posterior tubercular area of diencephalon was studied in four species of the cartilaginous ganoids by using routine Nissl staining and Bielschowski impregnation technique in Viktorov’s modification. The posterior tubercular area in the giant sturgeon Huso huso L., the Kura sturgeon Acipenser güldenstädtii persicus n. Kurensis Belyaeff, the stellate sturgeon Ac. stellatus Pall., and the barbel sturgeon Ac. nudiventris Lov. was shown to have similar structure. Six structures were identified in these areas: the periventricular nucleus, paraventricular organ, nucleus of the paraventricular organ, and posterior tuberal nucleus in the periventricular region and preglomerular medial and lateral nuclei in the rostral region. Both nuclei of the rostral zone are migrated nuclei. Out of nuclei of the posterior tubercle regions, the posterior tuberal and medial preglomerular nuclei are characterized by polymorphism of cellular elements. A conclusion is made that these parts of the posterior tubercular area in the sturgeons considered to be the lower ray-finned fish are comparable with those of the higher teleosts, and even are more differentiated according to some parameters.     

Processes of Proliferation and Apoptosis in the Brain of the Amur Sturgeon

Using techniques of immunoperoxidase staining of proliferating cell nuclear antigen (PCNA) and TUNEL labeling of fragmented DNA, we studied sites of proliferation and apoptosis in the myelencephalon, cerebellum, tectum opticum, thalamus, and hypothalamus of the Amur sturgeon (Acipenser schrenckii). We found that the processes of proliferation and apoptosis are maintained in the brain of 3-year-old sturgeon individuals; the ratio of these processes in different cerebral regions varied significantly. The maximum intensity of proliferative activity was found in the periventricular zone of the myelencephalon (proliferation index, on average, 21.0 ± 1.3%). This fact allows us to consider this cerebral region a most important zone were adult neurogenesis occurs in the sturgeon. In the medial reticular formation, dorsal thalamic nuclei, inner fibrous layer of the tectum, and lateral hypothalamus, the maximum numbers of apoptotic elements were found. Therefore, these zones in the brain of the sturgeon correspond, apparently, to the regions where postmitotic neuroblasts are localized. In sensory centers (tectum and nuclei of the V, VII, and X nerves), significantly varying ratios of intensities of proliferation and apoptosis were found; this is indicative of dissimilar rates of growth and differentiation in visual and chemosensory centers of the sturgeon brain. The high proliferative activity in sensory and motor cerebral centers of the sturgeon allows us to hypothesize that a neotenic pattern is preserved in these CNS regions of adult sturgeons over a long period after the embryogenesis has been completed.     

Formation of populations of Acipenseridae sturgeons in the Volga-Caspian basin

Analysis of dynamics of commercial catches and the state of populations of Russian sturgeon Acipenser gueldenstaedtii, starred sturgeon A. stellatus, and great sturgeon A. huso (Huso huso) in the Caspian Sea is presented. In the retrospective, the role of natural and artificial reproduction in formation of populations of Acipenseridae sturgeons is considered. The main principles of rational organization of practical activity for preservation and restoration of populations of Acipenseridae sturgeons are formulated.     

High‐throughput SNP‐genotyping analysis of the relationships among Ponto‐Caspian sturgeon species

Legally certified sturgeon fisheries require population protection and conservation methods, including DNA tests to identify the source of valuable sturgeon roe. However, the available genetic data are insufficient to distinguish between different sturgeon populations, and are even unable to distinguish between some species. We performed high‐throughput single‐nucleotide polymorphism (SNP)‐genotyping analysis on different populations of Russian (Acipenser gueldenstaedtii), Persian (A. persicus), and Siberian (A. baerii) sturgeon species from the Caspian Sea region (Volga and Ural Rivers), the Azov Sea, and two Siberian rivers. We found that Russian sturgeons from the Volga and Ural Rivers were essentially indistinguishable, but they differed from Russian sturgeons in the Azov Sea, and from Persian and Siberian sturgeons. We identified eight SNPs that were sufficient to distinguish these sturgeon populations with 80% confidence, and allowed the development of markers to distinguish sturgeon species. Finally, on the basis of our SNP data, we propose that the A. baerii‐like mitochondrial DNA found in some Russian sturgeons from the Caspian Sea arose via an introgression event during the Pleistocene glaciation.     

Organization of diencephalon of the sturgeons. Preoptic area

Cytoarchitectonics of preoptic area of diencephalon of chondroid ganoid fish (the sturgeons) was studied in serial sections using Nissl and Bilschowski techniques, the latter in Viktorov’s modification). The preoptic area of the hausen, Huso huso L., Kura sturgeon, Acipenser guldenstaedtii persicus n. kurensis Belyaeff, Caspian sturgeon, Acipenser stellatus Pall. and the sturgeon Acipenser nudioventris Lov. was shown to be one of the largest parts of diencephalon. It is located along the rostral excess of the third ventricle and the anterior third of the ventricle thalamic part. Eight nuclei are identified in the preoptic area: (1) anterior microcellular preoptic nucleus, (2) suprachiasmatic nucleus, (3) accessory preoptic nucleus, (4) intrachiasmatic nucleus, (5) magnocellular preoptic nucleus, (6) posterior microcellular preoptic nucleus, (7) caudal preoptic nucleus, (8) entopeduncular nucleus. Besides, microcellular, magnocellular, and gigantocellular parts are identified in the magnocellular preoptic nucleus. The obtained results indicate high differentiation of this area of the sturgeons among Actinopterygii.     

NADPH-Diaphorase-containing neurons in the medullary structures involved in generation of the respiratory activity in neonatal rats

Using a histochemical technique, we examined distribution of the neurons containing a marker of nitric oxide synthase (NOS), NADPH-diaphorase (NADPH-d), on frontal slices of the medulla and upper cervical spinal segments of 4-day-old rats. It was demonstrated that NADPH-d-positive cells are present within the dorsal and ventral medullary respiratory groups. The highest density of the labeled middle-size multipolar neurons (27.9±2.6 cells per 0.1 mm² of the slice) was observed in the rostral part of the ventral respiratory group, within the reticular lateral paragigantocellular nucleus. Similar NADPH-d-positive neurons were also observed in other reticular formation structures: rostroventrolateral reticular, gigantocellular, and ventral medullary nuclei, and in the ventral part of the paramedial nucleus. There were no labeled neurons in the lateral reticular nucleus. Single small and medium-size labeled neurons were found at all rostro-caudal levels of thenucl. ambiguous (nuclei retrofacialis, ambiguous, andretroam-biguous). Groups of NADPH-d-positive neurons were also revealed within the dorsal respiratory group, along the whole length of thenucl. tractus solitarii (mostly in its ventrolateral parts). Single labeled neurons were also observed in thenucl. n. hypoglossi, and their groups were observed in the dorsal motor part of thenucl. n. vagus. Involvement of the structures containing NADPH-d-positive neurons in the processes related to generation of the respiratory activity is discussed. Our neuroanatomical experiments prove that in early postnatal mammals NO is actively involved in generation and regulation of the medullary respiratory rhythm. Neirofiziologiya/Neurophysiology, Vol. 32, No. 2, pp. 128–136, March–April, 2000.     

The validation of specific status of the sakhalin sturgeon <Emphasis Type="Italic">Acipenser mikadoi</Emphasis> (Acipenseridae) in the light of recent genetic and morphological data

The comparative morphological study in different Acipenseriform species demonstrates that there are only two morphological features combining the kaluga and the great sturgeon and separating them from other sturgeons, namely the shape of a mouth and the manner attaching of gill membranes to isthmus in adult specimens; whereas many morphological characters are different in these species. The relations of morphological similarity/distinction among different sturgeon species and polyphyletic origin of genus Huso revealed by both molecular and cytogenetic studies presume the restoration of initial taxonomic states for the great sturgeon and kaluga as members of the same genus Acipenser, namely A. huso and A. dauricus.     

Distribution and quantitative characterization of NADPH-diaphorase-reactive neurons in analgesic zones of the rat midbrain

The distribution of nitric oxide synthase (nicotinamide adenine dinucleotide phosphate diaphorase, NADPH-d)-containing neurons in the rat midbrain was studied. We found that NADPH-d-reactive neurons were predominantly concentrated in the dorsolateral part of the periaqueductal gray (PAG) and the dorsal raphe nucleus, which are implicated in the control of nociceptive transmission. Such neurons were also present in the supraoculomotor cap and laterodorsal tegmental nuclei. In the dorsolateral part ofPAG, the moderately stained small fusiform cells were revealed. In the dorsal raphe nucleus and laterodorsal tegmental nuclei, the densely stained multipolar or oval cells of larger size dominanted. The NADPH-d-reactive cells were not found in the ventrolateral part of central gray, which is considered the main source of antinociceptive descending influences. Quantitative analysis of histochemically revealed neurons showed that their number is somewhat higher in the caudal parts of dorsolateral central gray and considerably higher in the rostral regions of some dorsal raphe subnuclei. This peculiarity was expressed in significant accumulation of the NADPH-d-reactive neurons at the midbrain levels from Fr –7.6 to –8.0. The possible involvement of the NO-synthase-containing class of neurons in the functional organization of analgesic zones and formation ofPAG antinociceptive output signals is discussed.Neirofiziologiya/Neurophysiology, Vol. 28, No. 1, pp. 36–46, January–February, 1996.     

Cytoarchitectonic pattern of the hypothalamus in the crocodile,Gavialis gangeticus

Summary The hypothalamus of the crocodile, Gavialis gangeticus, was investigated to reveal the organization of various nuclear complexes and to suggest homologies. The hypothalamic nuclei of G. gangeticus are composed of magnocellular and parvocellular neuronal entities. In the magnocellular system the nucleus supraopticus is well developed, whereas the nucleus paraventricularis and nucleus retrochiasmaticus are represented by scattered somata. Application of cytoarchitectonic criteria permits the delineation of 24 distinct parvocellular nuclear complexes extending rostrocaudally from the anterior commissure to the level indicated by the median eminence and nucleus mamillaris; some are further divisible into subgroups. The nucleus of the preoptic recess appears to be a unique property of the crocodilian hypothalamus. The nucleus suprachiasmaticus possesses a wing-like ventrolateral expansion that protrudes along the lateral aspect of the optic nerve. The tuberal region displays an elaborate pattern of nuclei segregated by regional specializations of the neuropil. The nucleus hypothalamicus posterior occupies the periventricular zone, flanked laterally by the nucleus hypothalamicus dorsomedialis and nucleus arcuatus. Further laterally, extended subdivisions of the nucleus hypothalamicus lateralis contain neurons rich in Nissl substance; the specializations shown by these subdivisions, in comparison to the lateral cell groups in lizards and snakes, are suggestive of enhanced integrative functions. The conspicuous paraventricular organ is encircled by dorsal and ventral divisions of the nucleus of the paraventricular organ. The neurons of the nucleus subfornicalis and nucleus hypothalamicus medialis are few in number, but large in size. The general organization of the hypothalamus of G. gangeticus reveals a mosaic-like pattern with the constituent groups appearing as clusters of small and large neurons, arranged medially and laterally in a definitive manner and accompanied by extensive zones of neuropil in the subependymal and lateral zones of the hypothalamus. The median eminence is divisible into an anterior and a posterior region. The nuclear pattern in the crocodilian hypothalamus reveals a higher state of morphologic organization compared to the situation in lizards or snakes, and thus reflects an evolutionary trend in the avian direction.     

Genetic monitoring of progeny derived from natural spawning of Acipenser ruthenus Linnaeus, 1758 and Acipenser stellatus Pallas, 1771 as well as their hybrids in the Lower Volga river during the period 2017 to 2019

The state of natural spawning of sturgeons in the Lower Volga River was investigated through genetic monitoring of juveniles (larvae, fingerlings, subadults) captured at seven traditionally used monitoring locations. Sampling was performed during the period 2017–2019. In total, 460 individuals were caught and genotyped using a set of six microsatellite loci (Afug41, Afug51, Afug135, An20, AoxD161, AoxD165) and mtDNA control region sequencing. The observed species-specific microsatellite alleles revealed the presence of juvenile sturgeons originating from natural spawning also included some hybrids between Acipenser stellatus and A. ruthenus. Thus, the majority of sterlet breeding takes place in natural spawning grounds but also sterlet hybridizes with a small number of stellate sturgeons. Juveniles of other sturgeon species were not identified, and it is concluded that natural spawning activities of Russian sturgeon, beluga and ship sturgeon were insignificant or even absent in the Lower Volga during the observation period.     

Features of inheritance of several morphological characters in the hybrids between Siberian sturgeon <Emphasis Type="Italic">Acipenser baerii</Emphasis> and beluga <Emphasis Type="Italic">A. huso</Emphasis> (Acipenseridae) and in gynogenetic individuals of Siberian sturgeon

Comparative analysis of several morphological characters is conducted in experimentally obtained hybrids between Siberian sturgeon Acipenser baerii and beluga A. huso, in parental species, and in gynogenetic progeny of Siberian sturgeon. The gynogenetic individuals are similar to Siberian sturgeon by all characters used for differentiation between Siberian sturgeon and beluga that supports gynogenetic origin of these individuals. Based on the data obtained on the hybrid specimens, the features of inheritance of the studied characters in sturgeons and their variation in postlarval ontogeny are discussed.     

Genetic variation and relationships of seven sturgeon species and ten interspecific hybrids

Background

Sturgeon cultivation is important for both industry and aquaculture in China. To date, more than 17 species or strains have been farmed for fillets and caviar production. Crossbreeding among different sturgeon species is frequent and the F2 hybrids are fertile. However, large-scale farming can have negative impacts on wild populations i.e. escape of exotic sturgeons and must be taken into consideration. Escape of exotic sturgeons can cause severe ecological problems, including threatening native sturgeon species once the exotic varieties become established or hybridize with native individuals. However, little is known about their genetic resources and variation.

Methods

Genetic diversity and introgression of seven sturgeon species were analyzed using mitochondrial DNA cytochrome oxidase subunit I (COI) and nine microsatellite markers. This study included 189 individuals from seven sturgeon species and 277 individuals from ten lineages of F2 hybrid strains.

Results

MtDNA COI sequences (632 bp long) were generated from 91 individuals across the 17 sturgeon strains and produced 23 different haplotypes. Haplotype diversity was high (h = 0.915 ± 0.015) and nucleotide diversity was low (π = 0.03680 ± 0.00153) in the seven sturgeon species and ten interspecific hybrids. Phylogenetic analyses resulted in almost identical tree topologies, and different haplotype structures were mainly related with sturgeons of different female parents. Analysis of molecular variance revealed that 81.73% of the genetic variance was due to matrilineal differences, while 9.40% resulted from strain variation. Pairwise Fst values obtained with POLYSAT software, were high among strains and ranged from 0.031 to 0.164. Admixture analysis assigned seven distinct groups and ten genotypes of admixed clusters composed of hybrid strains using STRUCTURE when assuming K = 7.

Conclusions

The interspecific mtDNA gene tree corresponded to the expected taxonomic divisions. These relationships were also supported by the results from the microsatellite analysis and contributed to unambiguously identify seven sturgeon species and ten F2 hybrid strains from sturgeon farms in China. Moreover, we found that introgressive hybridization is pervasive, exists in both purebred and hybrid sturgeons, and may reflect widespread mismanagement in sturgeon breeding in China.     

Estimation of gonad volume, fecundity, and reproductive stage of shovelnose sturgeon using sonography and endoscopy with application to the endangered pallid sturgeon

Most species of sturgeon are declining in the Mississippi River Basin of North America including pallid (Scaphirhynchus albus F. and R.) and shovelnose sturgeons (S. platorynchus R.). Understanding the reproductive cycle of sturgeon in the Mississippi River Basin is important in evaluating the status and viability of sturgeon populations. We used non‐invasive, non‐lethal methods for examining internal reproductive organs of shovelnose and pallid sturgeon. We used an ultrasound to measure egg diameter, fecundity, and gonad volume; endoscope was used to visually examine the gonad. We found the ultrasound to accurately measure the gonad volume, but it underestimated egg diameter by 52%. After correcting for the measurement error, the ultrasound accurately measured the gonad volume but it was higher than the true gonad volume for stages I and II. The ultrasound underestimated the fecundity of shovelnose sturgeon by 5%. The ultrasound fecundity was lower than the true fecundity for stage III and during August. Using the endoscope, we viewed seven different egg color categories. Using a model selection procedure, the presence of four egg categories correctly predicted the reproductive stage ± one reproductive stage of shovelnose sturgeon 95% of the time. For pallid sturgeon, the ultrasound overestimated the density of eggs by 49% and the endoscope was able to view eggs in 50% of the pallid sturgeon. Individually, the ultrasound and endoscope can be used to assess certain reproductive characteristics in sturgeon. The use of both methods at the same time can be complementary depending on the parameter measured. These methods can be used to track gonad characteristics, including measuring Gonadosomatic Index in individuals and/or populations through time, which can be very useful when associating gonad characteristics with environmental spawning triggers or with repeated examinations of individual fish throughout the reproductive cycle.