Effect of photoperiod on pineal gland volume and pinealocyte size in the Chinese hamster, Cricetulus griseus

Male adult (200-day-old) Chinese hamsters (Cricetulus griseus) raised from weaning under either LD 16:8 or LD 8:16 were used. The pineal gland of the Chinese hamster consists of superficial (major) and deep (minor) components and a continuous, or interrupted, narrow parenchymal stalk interposed between them. The volume of the superficial pineal including the parenchymal stalk is greater under LD 16:8 than under LD 8:16. Under both photoperiods, pinealocytes in the superficial pineal have larger nuclei and more abundant cytoplasm than those in the deep pineal. Nuclei in the superficial pineal appear pale and usually have irregular profiles, whereas those in the deep pineal appear dark and have round profiles. In the superficial pineal, pinealocyte nuclei are larger, paler, and more irregular; and, in addition, nuclear density is lower under LD 16:8 than under LD 8:16. Similar, but less prominent, photoperiod-induced changes occur in the volume of the deep pineal, the size of pinealocytes, and pinealocyte nuclear morphology in the deep pineal. The results indicate that the development and differentiation of pinealocytes in both pineal portions may be advanced under long photoperiods and delayed under short photoperiods, although pinealocytes in the deep pineal may remain not fully differentiated even in adults. Since testicular weights and body weights are similar under both photoperiods, the photoperiod may exert marked influences on the development of the pineal gland without affecting reproductive activity and growth rates of animals.     

Differential permeability of pineal capillaries to lanthanum ion in the rat (Rattus norvegicus), gerbil (Meriones unguiculatus) and golden hamster (Mesocricetus auratus)

Summary Capillaries in the pineal gland of the rat (Rattus norvegicus), gerbil (Meriones unguiculatus) and golden hamster (Mesocricetus auratus) were investigated by means of electron-microscopical tracer studies using lanthanum. The tracer was administered together with the fixative solution by perfusion via the left cardiac ventricle. In the rat, endothelial junctions of fenestrated capillaries are permeable to lanthanum. In the gerbil the tracer does not leave the capillaries, which are of the non-fenestrated type throughout the organ. In the golden hamster the two portions of the pineal system have different types of capillaries. While in the superficial pineal fenestrated capillaries permeable to lanthanum predominate, the deep pineal possesses capillaries that are, without exception, devoid of fenestrations and impermeable to the tracer. It is suggested that differences in the structure of the capillaries are related to differences in the extent of a specialized contact of the pineal gland to the third ventricle.The authors dedicate this paper to Professor H. Rollhäuser, Münster, on the occasion of his 65th birthday     

Central innervation of the pineal organ of the Mongolian gerbil

Nerve fibers connecting the brain with the pineal gland of the Mongolian gerbil (central pinealopetal fibers) were investigated by means of light and electron microscopy. Several myelinated fibers penetrate from the brain into the deep pineal gland, extend further into the pineal stalk and continue to the superficial portion of the pineal gland. In the centripetal direction these fibers were traced to the stria medullaris and to the habenular nuclei, where they turned laterad and then occupied a position immediately ventral to the optic tract. As shown in electron micrographs, lesions of the habenular area led to degeneration of myelinated fibers and nerve boutons in the deep pineal gland, the pineal stalk and the superficial pineal gland. Only boutons containing clear transmitter vesicles (devoid of a dense core) were observed to degenerate after the habenular lesions. On the other hand, removal of the superior cervical ganglia resulted in degeneration of boutons containing small (40 to 60 nm in diameter) dense-core vesicles. Several of the nerve fibers that penetrate into the deep pineal directly from the brain (central fibers) exhibited a positive reaction for acetylcholinesterase (AChE). AChE-positive perikarya were located in the projections of the stria medullaris, the lateral portions of the deep pineal, the area of the posterior commissure, and the periventricular gray of the mesencephalon. Such perikarya were found neither in the pineal stalk nor in the superficial pineal gland. These results present anatomical evidence that the pineal organ of the Mongolian gerbil receives multiple nervous inputs mediated by peripheral autonomic (i.e., sympathetic) nerve fibers, on the one hand, and by central fibers, on the other.     

黑线毛足鼠和金仓鼠血清IgG的纯化及抗血清的制备

目的 纯化黑线毛足鼠和金仓鼠血清IgG,制备兔抗金仓鼠和黑线毛足鼠血清IgG的抗血清。方法 用Hitrap Protein G亲和层析纯化黑线毛足鼠和金仓鼠血清IgG,经SDS-PAGE电泳鉴定纯度,标准免疫方法免疫兔子制备抗血清。结果 黑线毛足鼠和金仓鼠血清IgG对protein G有很高的亲合性,用Hitrap Protein G亲和层析纯化,得到高纯度黑线毛足鼠和金仓鼠IgG,利用纯化的IgG作抗原制备了高效价的抗血清,免疫双扩散测定效价达1∶32和1∶16。结论 证实黑线毛足鼠和金仓鼠IgG和Protein G具有很高的亲和性,Protein G亲和层析是纯化黑线毛足鼠和金仓鼠IgG有效的方法之一,制备了黑线毛足鼠和金仓鼠IgG的抗血清。     

Indications for the presence of two populations of serotonin-containing pinealocytes in the pineal complex of the golden hamster (Mesocricetus auratus)

Summary Serotonin-like immunoreactivity was investigated in the pineal complex of the golden hamster by use of the indirect immunohistochemical technique. The superficial and deep portions of the pineal gland, and also the pineal stalk exhibited an intense cellular immunoreaction for serotonin. In addition, perivascular serotonin-immunoreactive nerve fibers were observed. Some serotonin-immunoreactive processes of the pinealocytes terminated on the surface of the ventricular lumen in the pineal and suprapineal recesses, indicating a receptive or secretory function of these cells. Several serotonin-immunoreactive processes connected the deep pineal with the habenular area. One week after bilateral removal of both superior cervical ganglia the serotonin immunoreaction of the entire pineal complex was greatly decreased. However, some cells in the pineal complex, of which several exhibited a neuron-like morphology, remained intensively stained after ganglionectomy. This indicates that the indoleamine content of some cells in the pineal complex of the golden hamster is independent of the sympathetic innervation.Supported by a Grant from the Italian Society for Veterinary Sciences     

Cytochemical studies on cytoplasmic granular elements in the hamster pineal gland.

The pineal gland of adult golden hamsters (Mesocricetus auratus) was studied by various cytochemical methods at the electron microscopic level: (1) the modified chromaffin reaction specific for 5-hydroxytryptamine (5-HT), (2) argentaffin reaction, (3) zinc-iodide-osmium (ZIO) mixture reaction and (4) acid phosphatase reaction. In the pinealocytes, the dense-cored vesicles (80-160 nm in diameter) show both chromaffinity and argentaffinity, while the population of dense bodies (150-400 nm in diameter) is reactive to ammoniacal silver solution and ZIO mixture but not to the modified chromaffin reaction. After incubation for demonstration of acid phosphatase activity, reaction products are localized in some, but not all, of the dense bodies, in some of the small vesicles in the Golgi region and in one or two inner Golgi saccules. In nerve fibers in the pineal gland, small granulated vesicles are also reactive to the modified chromaffin reaction and ZIO mixture. Based upon these cytochemical results the following conclusions have been reached: (1) dense cored vesicles in the pinealocytes and small granulated vesicles in the nerve fibers of the hamster pineal gland contain 5-HT, and (2) the population of dense bodies in the pinealocytes is heterogeneous, some are lysosomes and the other are possibly the granules responsible for the secretion of pineal peptides.     

金黄地鼠与白化地鼠遗传生化基因位点概貌

目的建立金黄地鼠和白化地鼠遗传生化基因位点。方法选用小鼠和大鼠的遗传生化基因位点,采用蛋白质和同工酶醋酸纤维电泳的方法,对金黄地鼠和白化地鼠进行生化基因位点检测。结果建立了金黄地鼠和白化地鼠25个生化基因位点,分析金黄地鼠和白化地鼠遗传生化基因位点的多态性,为进一步研究金黄地鼠白化突变系的遗传机理奠定基础。结论金黄地鼠生化基因位点存在多态性,白化地鼠与金黄地鼠生化基因位点存在差异。     

Excitatory amino acid receptors may mediate the effects of light on the reproductive system of the golden hamster.

Seasonal regulation of reproduction occurs in golden hamsters (Mesocricetus auratus) in response to changes in day length (photoperiod). We report here the results of experiments designed to evaluate whether excitatory amino acid neurotransmission is involved in mediating the effects of light on reproduction. We found that MK-801, a specific antagonist of the N-methyl-D-aspartate subclass of excitatory amino acid receptors, blocked the stimulatory action of short pulses of light on gonadal function in hamsters maintained in short days. Furthermore, MK-801 also inhibited the light-induced decline in pineal melatonin content. These findings suggest that excitatory amino acids may mediate the effects of light on the reproductive system.     

Pineal melatonin in hibernating and aroused golden hamsters (Mesocricetus auratus)

Daily changes of pineal melatonin content were determined in warm-adapted nonhibernating and cold-adapted hibernating golden hamsters (Mesocricetus auratus). Pineal melatonin in nonhibernating golden hamsters showed marked daily rhythm with the night values about 20 times higher than the daytime ones. In hamsters hibernating for 2 and 3 days the melatonin rhythm was abolished, since no increase of pineal melatonin over basal levels occurred throughout 24 hr period. After arousal from hibernation melatonin increased rapidly regardless whether the hamsters were provoked to arousal during day or night.     

Morphometric analysis of the pineal complex of the golden hamster over a 24-hour light:dark cycle: I. The superficial pineal in untreated and optically enucleated animals

Morphometric analysis of the superficial pineal gland of intact and blinded golden hamsters was conducted at both the light and electron microscopic level. The volume of the superficial gland was estimated to be 151 X 10(6) micron 3, comprising 90-94% of the total pineal parenchymal tissue. Analysis of structural rhythms in animals maintained under a 14:10 L:D cycle showed significant 24-hr variations in values for pinealocyte nuclei, nucleoli, rough and smooth endoplasmic reticulum, Golgi bodies, dense bodies, and dense-cored vesicles. Peak values for these structures generally occurred at the light:dark interface. These results provide morphological correlates for known rhythmic variations in the synthesis of pineal-gland products. Superficial pineals examined 8 weeks following optic enucleation exhibited a decrease in the volume of pinealocyte nuclei and cytoplasm, while nucleolar size and the amounts of smooth and rough endoplasmic reticulum, Golgi bodies, dense bodies and dense-cored vesicles were enhanced. The latter changes are interpreted as indications of increased synthetic activity by the superficial pineal gland in response to light deprivation.     

Morphometric analysis of the pineal complex of the golden hamster over a 24-hour light:dark cycle: II. The deep pineal in untreated and optically enucleated animals

The deep pineal gland of golden hamsters was morphometrically analyzed and quantitatively compared with the superficial pineal under a 14:10 lighting regime and following blinding. The deep pineal comprised 6-10% of the total pineal parenchymal tissue. Pinealocytes of the deep gland were smaller than the cells of the superficial pineal and showed a greater percent volume of Golgi bodies, rough endoplasmic reticulum, and dense-cored vesicles. Twenty-four-hour rhythms in nucleoli and Golgi bodies were found in deep pinealocytes. These rhythms were out of phase with comparable rhythms in the superficial pineal gland, suggesting that distinct subpopulations of pinealocytes are present within the respective parts. Blinding resulted in decreased nuclear and nucleolar volume, while the amount of smooth endoplasmic reticulum, Golgi bodies, dense bodies, and dense-cored vesicles increased significantly. Marginal increases were seen in mitochondria and lipid droplets. The greater abundance of those organelles involved in synthesis and secretion suggests enhanced cellular activity after blinding. Many of the morphological responses are similar to alterations in the pinealocytes of the superficial pineal following optic enucleation.     

Photoperiodic modulation of sexual and aggressive behavior in female golden hamsters (Mesocricetus auratus): role of the pineal gland

Pinealectomized female hamsters (Mesocricetus auratus) housed in a short-day photoperiod were ovariectomized and tested for hormone-induced sexual receptivity in order to investigate the role of the pineal gland in the control of behavioral sensitivity to exogenous ovarian steroid hormones (Experiment 1). Behavioral sensitivity to hormones was further investigated in females maintained in a long-day photoperiod and rendered acyclic by daily administration of exogenous melatonin (Experiment 2). Female aggressive behavior was also monitored in all tests. Pinealectomy did not affect the reduced behavioral sensitivity to exogenous estrogen (E) induced by short days. These animals were also partially refractory to the effects of E when combined with low doses of progesterone. In addition, although melatonin administration mimicked the effects of short days on estrous cyclicity, the expression of hormone-dependent behaviors in these animals resembled the pattern displayed by control animals kept in long days. Thus, these findings suggest that the pineal gland plays a negligible role in the photoperiodic modulation of hormone-dependent sociosexual behaviors in female hamsters.     

Internal architecture, origin-insertion site, and mass of jaw muscles in Old World hamsters

The jaw muscle (i.e., masticatory, suprahyoid, and extrinsic tongue) anatomy and mass were examined in four genera of Old World hamsters (cricetine murids), Mesocricetus, Cricetulus, Tscherskia, and Phodopus. The masseter was the largest and most complicated of the muscles examined. In the superficial layer, a few ventral fibers form a small medially turned portion with an insertion site more similar to those of sciurids than of other murids. In Mesocricetus, the superficial layer has a discrete anteroventral portion that has not been reported for other murid rodents. Examination of the fiber attachment sites indicated that the deep layer contains four parts and the medial layer contains three parts. The deep layer originates from two aponeuroses that are firmly connected to each other at their anterior ends and lie along the zygomatic arch. The aponeurosis of insertion for the deep layer is situated along the masseteric ridge and the dorsal border of the angular process, but is absent in its middle part, consistent with reports in two relatives, sigmodontine and arvicoline murids. In cricetine murids, unlike in other rodents, fibers insert on the dorsal narrow strip of the posterior mandibular aponeurosis, not on its broad medial aspect. The relative mass of some masticatory and suprahyoid muscles is related to body mass. Small species (Cricetulus and Phodopus) have relatively larger masseter and mylohyoid muscles and smaller temporalis and geniohyoid muscles than large species (Mesocricetus and Tscherskia).     

Morphological investigation of the deep pineal of the rat

Summary The results presented here reveal that in adult Sprague-Dawley and Wistar rats the pineal gland represents a complex rather than a single organ. Regularly one can distinguish (i) pineal tissue in the intercommissural region as a deep pineal, (ii) a superficial pineal, which represents the major part of the pineal complex, and (iii) nearly always a parenchymal stalk of variable length. The volume of the deep pineal with the adjacent parenchymal stalk exhibits great interindividual variation. It amounts to 127±39×10⁵ m³ (mean ±standard deviation). The histological appearance of the deep and superficial pineal tissue is fairly similar. The intrinsic cells of the deep and superficial pineal differ in nuances only. Karyometry reveals that the nuclear volumes of the intrinsic cells of the deep pineal are very variable ranging from 90–450 m³, with a mean value of 207 m³. The changes over a period of 24 h reach statistical significance.Supported by the Deutsche Forschungsgemeinschaft (Grant Vo 135/4) within the Schwerpunktprogramm NeuroendokrinologieThis paper is an abridged version of a thesis submitted for obtaining the degree of Dr. med., Fachbereich Medizin, University of Mainz     

Effect of superior cervical ganglionectomy on monoamine content in the epithalamic area of the Mongolian gerbil (Meriones unguiculatus): A fluorescence histochemical study

Summary Extirpation of the superior cervical ganglion was performed in a series of Mongolian gerbils. One or two weeks after the ganglionectomy the animals were injected with a monoamine oxidase inhibitor. Subsequently perfusion fixation was performed using the glyoxylic acid-paraformal-dehydemagnesium method (Lorén et al., 1976) for fluorescence histochemical investigation of the monoamines of the pineal complex. In the ganglionectomized animals all of the blue-fluorescent sympathetic fibers in the pineal complex (superficial pineal gland, deep pineal gland and the pineal stalk) completely disappeared. The yellow indolamine fluorescence of the cells in the superficial pineal and the deep pineal, as well as in the pineal stalk, was markedly reduced after ganglionectomy. No change in the morphology or number of sympathetic fibers in the medial habenular nucleus was observed. These results indicate that the presence of sympathetic nerve fibers with perikarya in the superior cervical ganglion is necessary for maintaining a high indolamine content in all three parts of the pineal complex. In addition, the results also indicate that the deep pineal gland is a functional part of the pineal complex. The presence of a functionally active deep pineal, bordering the pineal recess, suggests that part of the pineal hormones might be secreted into the cerebrospinal fluid.This work was supported by the Carlsberg Foundation, the Swedish Natural Science Research Council, grant no. 2126-100, and the Danish Medical Research Council, grant no. 512-7134     

Tyrosine hydroxylase- and neuropeptide Y-immunoreactive nerve fibers in the pineal complex of untreated rats and rats following removal of the superior cervical ganglia

Summary The distribution of tyrosine hydroxylase (TH)- and neuropeptide Y (NPY)-immunoreactive(IR) nerve fibers in the pineal complex was investigated in untreated rats and rats following bilateral removal of the superior cervical ganglia. In normal animals, a large number of TH- and NPY-IR nerve fibers were present in the pineal capsule, the perivascular spaces, and intraparenchymally between the pinealocytes throughout the superficial pineal and deep pineal gland. A small number of TH-IR and NPY-IR nerve fibers were found in the posterior and habenular commissures, a few fibers penetrating from the commissures into the deep pineal gland. To elucidate the origin of these fibers, the superior cervical ganglion was removed bilaterally in 10 animals, and the pineal complex was examined immunohistochemically. Two weeks after the ganglionectomy, the TH-IR and NPY-IR nerve fibers in the superficial pineal gland had almost completely disappeared. On the other hand, in the deep pineal and the pineal stalk, the TH-IR and NPY-IR fibers were still present after ganglionectomy. These data show that the deep pineal gland and the pineal stalk possess an extrasympathetic innervation by TH-IR and NPY-IR fibers. It is suggested that the extrasympathetic TH-IR and NPY-IR nerve fibers innervating the deep pineal and the pineal stalk originate from the brain.     

A histochemical study of aldehyde fuchsin-positive material and "high-esterase cells" in the pineal gland of the Mongolian gerbil.

The pineal gland of the Mongolian gerbil consists of a superficial gland, stalk and deep pineal. The deep pineal differentiates postnatally. Histochemical studies of the superficial pineal gland indicate that it may be involved in the secretion of protein. Presumptive secretory material visualized by aldehyde fuchsin (AF) and chrome hematoxylin was observed along the course of blood vessels and among the pinealocytes. The distribution and texture of the AF-positive material was distinctive. It did not correspond to the pattern and texture of material stained with PAS, Sudan Black or acid orcein. Staining with AF was markedly reduced after incubation with trypsin, indicating that the AF-positive material is at least partially protein. The amount of stainable material increased with age. The AF-positive material was observed in what appeared to be interstitial or glial cells and processes, and in the processes of perivascular cells. Cells and fibrous processes with high non-specific esterase activity ("high-esterase cells") were observed among the pinealocytes and along the course of blood vessels. The distribution of the "high-esterase cells" and the morphology and texture of their esterase-containing processes were remarkably similar to the morphology and distribution of the material that stained with AF. It may be that the "high-esterase cells" contain AF-positive material. The "high-esterase cells" hydrolyzed both alpha-naphthyl acetate and alpha-naphthyl butyrate. The pinealocytes hydrolyzed only alpha-naphthyl acetate. The "high-esterase cells" appear to form a distinct class of cells within the superficial pineal gland. They are tentatively identified as a type of glial cell.     

雌雄金黄地鼠血液生理生化指标的比较分析

目的测定金黄地鼠的血液生理生化参考值,并对两性的参考值进行比较。方法将88只4周龄金黄地鼠,80只8周龄金黄地鼠,采血后,用日本光电MEK-7222K血球分析仪测定血液生理指标,用Olympus AU400全自动生化分析仪测定血清生化指标。结果4周龄金黄地鼠血液生理指标中RBC、HGB、HCT、MCV和RDW性别差异有显著性（P〈0.05）,而WBC、MCH、MCHC、PLT、PCT、MPV和PDW等性别差异无显著性（P〉0.05）;4周龄金黄地鼠血清生化指标中ALB、ALP、GLU、CA、P、CHO和TG性别差异有显著性（P〈0.05）,而ALT、AST、TP、BUN和CREA等性别差异无显著性（P〉0.05）。8周龄金黄地鼠血液生理指标中WBC、LYM、MON、BAS、LYM％、NEUT％、MCH、RBC、HGB、HCT、MCV、PDW和RDW性别差异有显著性（P〈0.05）,而MCHC、PLT、MPV、PCT、NEUT、EOS、BAS、MON％和EOS％等性别差异无显著性（P〉0.05）;8周金黄地鼠血清生化指标中ALT、ALB、BUN、GLU、CREA、P、CHO和TG性别差异有显著性（P〈0.05）,而AST、TP、ALP和CA等性别差异无显著性（P〉0.05）。结论金黄地鼠血液的生理生化值受多种因素影响,部分指标两性问测定值有差异,但多数指标差异不显著。     

中国地鼠线粒体Cyt b基因测序及其分子进化

目的测定中国地鼠线粒体DNA细胞色素b基因部分序列,分析其分子系统进化关系。方法提取中国地鼠肝脏的总基因组DNA。设计合成特异引物进行PCR扩增,经检测进行测序。用Blast与GenBank中啮齿类其他常用实验动物的物种细胞色素b基因进行同源序列比较,分析其碱基组成及变异情况,并用邻接法、最大简约法、最小进化法构建了分子系统树,在分子水平上探讨中国地鼠和常用啮齿类实验动物的进化关系。结果获得了中国地鼠线粒体Cytb基因的部分序列,共936bp。结论中国地鼠和金黄地鼠的亲缘关系最近,与小鼠、大鼠存在的差异相对大,与豚鼠的亲缘关系最远,与传统的分类地位基本吻合。