Distribution of ghrelin-like immunoreactive cells in amphioxus, Branchiostoma belcheri – A study of immunohistochemistry

The distribution of ghrelin-like immunoreactive cells in amphioxus (Branchiostoma belcheri) was investigated by using immunohistochemical staining with rabbit antiserum against synthetical mammalian ghrelin. The results showed that ghrelin-like immunoreactive cells were distributed widely in the nervous system, Hatschek’s pit, wheel organ, digestive tract and gonads (ovary and testis). In nervous system, ghrelin-like immunoreactive neurons and their protrusions were distributed speci?cally on the dorsal side, ventral side and funnel part of brain vesicle, with a few dispersive immunoreactive nerve cells and their ?bers in nerve tube. Ghrelin-like immunoreactivities were also detected in Hatschek’s pit epithelial cells and wheel organ cells, with positive substance located along cell membrane. In digestive tract, ghrelin-like immunoreactive cells existed in hepatic diverticulum, anterior and posterior region of midgut, and could be classified into two types, closed- and opened-type endocrine cells. The number of positive cells was most in hepatic diverticulum, secondary in posterior region of midgut and least in anterior region of midgut. In gonads, ghrelin-like immunoreactive substance was detected in oogonia, oocytes and follicle cells in ovary at the small and large growth stages and in early spermatogenic cells and Sertoli cells in testis. The extensive distribution of ghrelin-like cells in amphioxus suggested that these kinds of cells are conservative in evolution and diversified in function. At the same time, we found for the ?rst time that ghrelin-like immunoreactive cells existed in brain vesicle and Hatschek’s pit, which provided new morphological evidence for the existence of an activation pathway between brain vesicle and Hatschek’s pit for the regulation of growth hormone excretion.     

Distribution of a-MSH-like immunoreactive cells in the nervous system,Hatschek's pit and other tissues of amphioxus,Branchiostoma belcheri

Immunohistochemical localization of α-melanocyte-stimulating hormone （α-MSH） in the nervous system, Hatschek＇s pit and other tissues of amphioxus （Branchiostorna belcheri） was performed using the antibody against synthetic α-MSH. The results revealed that α-MSH-like immunoreactive cells were distributed at the dorsal side and ventral side of brain vesicle, the dorsal side and the surrounding of nerve tube, and in the epithelial cells of Hatschek＇s pit, the zone 1, 3, and 6 of endostyle and gut. The immunoreactive substance was also found in the primary oocytes of the small and large growth stage of ovary and early stage spermatogenic cells in testis. These findings indicate that α-MSH is an ancient and highly conserved hormone and it is extensively distributed in amphioxus. Although Hatschek＇s pit in amphioxus does not have a structure of the intermediate lobe of vertebrate adenohypophysis, it has already hosted α-MSH-like endocrine cells, implying that the functional differentiation of α-MSH-like cells occurred earlier than the differentiation of the tissue structure. The results of the present study provided a new evidence for the endocrinology of Hatschek＇s pit and for the origin and evolution of vertebrate adenohypophysis.     

Distribution of growth hormone-like immunoreactive cells and somatostatin receptors in the nervous system and Hatschek''s pit of amphioxus, Branchiostoma belcheri

Using immunohistochemical method and double staining technique, the localization of growth hormone (GH) and somatostatin receptors in the nervous system and Hatschek's pit of amphioxus has been investigated. The results showed that the growth hormone-like nerve cells and endocrine cells as well as three subtypes of somatostatin receptors exist in the nervous system and Hatschek's pit, and GH-like nerve cells and endocrine cells co-exist with three subtypes of somatostatin receptors in the brain vesicle and Hatschek's pit. It is suggested that a primitive control system of inhibitory growth hormone secretion in Hatschek's pit could have been developed in amphioxus, as in vertebrates. The present study provides new evidence for the endocrinology and the evolution of Hatschek's pit.     

Distribution of growth hormone-like immunoreactive cells and somatostatin receptors in the nervous system and Hatschek's pit of amphioxus, Branchiostoma belcheri

Using immunohistochemical method and double staining technique, the localization of growth hormone (GH) and somatostatin receptors in the nervous system and Hatschek's pit of amphioxus has been investigated. The results showed that the growth hormone-like nerve cells and endocrine cells as well as three subtypes of somatostatin receptors exist in the nervous system and Hatschek's pit, and GH-like nerve cells and endocrine cells co-exist with three subtypes of somatostatin receptors in the brain vesicle and Hatschek's pit. It is suggested that a primitive control system of inhibitory growth hormone secretion in Hatschek's pit could have been developed in amphioxus, as in vertebrates. The present study provides new evidence for the endocrinology and the evolution of Hatschek's pit.     

Distribution of α-MSH -like immunoreactive cells in the nervous system, Hatschek’s pit and other tissues of amphioxus, Branchiostoma belcheri

Immunohistochemical localization of α-melanocyte-stimulating hormone (α-MSH) in the nervous system, Hatschek’s pit and other tissues of amphioxus (Branchiostoma belcheri) was performed using the antibody against synthetic α-MSH. The results revealed that α-MSH-like immunoreactive cells were distributed at the dorsal side and ventral side of the brain vesicle, the dorsal side and the surrounding of central tube in the nerve tube, the epithelial cells of Hatschek’s pit, and zones 1, 3 and 6 in endostyle and hindgut. The immunoreactive substance was also found in the primary oocytes of the small and large growth stage of ovary and early stage spermatogenic cells in testis. These findings indicate that α-MSH is an ancient and highly conserved hormone and it is extensively distributed in amphioxus. Although Hatschek’s pit in amphioxus do not have a structure of the intermediate lobe of vertebrate adenohypophysis, it has already hosted α-MSH-like endocrine cells, implying that the functional differentiation of α-MSH-like cells occurred earlier than the differentiation of the tissue structure. The results of present study provided new evidence for the endocrinology of Hatschek’s pit and the origin and evolution of vertebrate adenphypophysis.     

Distribution of CRH-and ACTH-like immunoreactive cells in amphioxus,Branchiostoma belcheri

Immunocytochemical studies on the nervous system,Hatschek's pit,digestive tract and gonads tissues of an amphioxus(Branchiostoma belcheri)were performed using polyclonal antibodies against human corticotrophin-releasing hormone(CRH)and human adrenocorticotropin(ACTH).The results showed that many CRH-like immunoreactive neurons were distributed specifically on the dorsal side and ventral side of brain vesicle,while a few CRH-like neurons and their fibers in spinal cord.At the same time,the epithelial cells in the basic region of Hatschek's pit were shown immunopositive to CRH antibody.In gonads(ovary and testis),CRH-immunopositive substance was localized in the cytoplasm near oocyte nucleus and in early spermatogenic cells.ACTH-like immunoreactivities were observed specially in the neurons and their protrusions localized on the ventral side of the brain vesicle and in spinal cord,and also in epithelial cells of Hatschek's pit,enteric neurons of digestive tract,oocytes in ovary and in early spermatogenic cells as well.It was found for the first time that CRH-like neurons existed in the middle region of brain vesicle(corresponding to the hypothalamus of vertebrates)and ACTH-like immunopositive cells existed in Hatschek's pit,implying that a control mechanism between brain vesicle and Hatschek's pit maybe had been already built in amphioxus as that in vertebrates.The present study will provide new morphological evidence for the origin and evolution of ACTH.In addition,the immunoreactivities of CRH and ACTH in the digestive tract and gonads suggested other physiological function of CRH and ACTH in amphioxus.     

促生长激素样免疫活性细胞和生长抑素受体在文昌鱼（Branchiostoma belcheri）神经系统和哈氏窝的分布

用免疫组织化学与双标记染色技术对文昌鱼神经系统和哈氏窝进行生长激素（GH）和生长抑素受体（SSTR）免疫组织化学定位.研究结果显示，神经系统和哈氏窝均存在生长激素样免疫活性神经细胞和内分泌细胞及生长抑素3种亚型受体，且脑泡GH样免疫活性神经细胞和哈氏窝GH样细胞均与3种亚型生长抑素受体共存.这些结果证明了文昌鱼可能建立类似脊椎动物原始的抑制哈氏窝分泌生长激素的调控系统，为哈氏窝内分泌学及其进化提供了新的证据.     

Distribution of growth hormone-like immunoreactive cells and somatostatin receptors in the nervous system and Hatschek''''s pit of amphioxus, Branchiostoma belcheri

Using immunohistochemical method and double staining technique, the localization of growth hormone (GH) and somatostatin receptors in the nervous system and Hatschek's pit of amphioxus has been investigated. The results showed that the growth hormone-like nerve cells and endocrine cells as well as three subtypes of somatostatin receptors exist in the nervous system and Hatschek s pit, and GH-like nerve cells and endocrine cells co-exist with three subtypes of somatostatin receptors in the brain vesicle and Hatschek s pit. It is suggested that a primitive control system of inhibitory growth hormone secretion in Hatschek s pit could have been developed in amphioxus, as in vertebrates. The present study provides new evidence for the endocrinology and the evolution of Hatschek's pit.     

Distribution Pattern of Plant Populations of Rhizophora stylosa Community in Yingluo Bay of Guangxi

This paper deals with the distribution pattern type,aggregative intensity,pattern scaleand dynamics of plant populations of Rhizophora stylosa community in Yingluo Bay of Guangxi.The measured results show that R.stylosa population is an aggregated distribution,while thepopulations of Kandelia candel,Bruguiera gymnorrhiza and Aegiceras corniculatum appear asrandom distribution.The plant population distribution pattern changes,along with the populationdevelopment and the succession of the community.     

A restudy of Oryctocephalus indicus (Reed, 1910)

Restudy on morphology of Oryctocephalus indicus (Reed, 1910), based on specimens from Guizhou, China and Nevada USA, suggests that the subspecies regarded previously as Oryctocephalus indicus indicus (Reed, 1910), O. indicus latus Zhao et Yuan 2002, and O. indicus kobayashi Saito, 1934, are synonymous, and Oryctocephalus americanus Sundberg et McCollum, 2003 is a similar form of O. indicus. Oryctocephalus indicus is rediagnosed as having a glabella that is subconical in outline and tapering forward slightly; a thorax that comprises 12 segments, and a small pygidium that bears 2?3 axial rings with a terminal piece and a postaxial ridge. Oryctocephalus indicus is widely distributed in eastern Guizhou and ranges through a great interval of Kaili Formation. Its first appearance is almost identical with these events of trilobite extinction-recovery occurring at the end of the Early Cambrian, the alternation of acritach assemblages, the change of trace elements (REE) and stable isotopes (carbon) at the Wuliu-Zengjiaya section. As one of the key forms for defining the traditional Lower - Middle Cambrian boundary (the base of Cambrian Series 3 and Stage 5 in the new Cambrian chronstratigraphic standard), O. indicus has received detailed study and has more advantages than other species such as Ovatoryctocara granulata Tchernysheva, 1962 or Arthricocephalus chauveaui Bergeron, 1899.     

Distribution of α-MSH-like immunoreactive cells in the nervous system, Hatschek''''s pit and other tissues of amphioxus, Branchiostoma belcheri

Immunohistochemical localization of a-melanocyte-stimulating hormone (α-MSH) in the nervous system, Hatschek's pit and other tissues of amphioxus (Branchiostoma belcheri) was performed using the antibody against synthetic α-MSH. The results revealed that α-MSH-like immunoreactive cells were distributed at the dorsal side and ventral side of brain vesicle, the dorsal side and the surrounding of nerve tube, and in the epithelial cells of Hatschek's pit, the zone 1, 3, and 6 of endostyle and gut. The immunoreactive substance was also found in the primary oocytes of the small and large growth stage of ovary and early stage spermatogenic cells in testis. These findings indicate that α-MSH is an ancient and highly conserved hormone and it is extensively distributed in amphioxus. Although Hatschek's pit in amphioxus does not have a structure of the intermediate lobe of vertebrate adenohypophysis, it has already hosted α-MSH-like endocrine cells, implying that the functional differentiation of α-MSH-like cells occurred earlier than the differentiation of the tissue structure. The results of the present study provided a new evidence for the endocrinology of Hatschek's pit and for the origin and evolution of vertebrate adenohypophysis.     

Distribution of CRH- and ACTH-like immunoreactive cells in amphioxus, Branchiostoma belcheri

Immunocytochemical studies on the nervous system, Hatschek’s pit，digestive tract and gonads tissues of an amphioxus (Branchiostoma belcheri) were performed using polyclonal antibodies against human corticotrophin-releasing hormone (CRH) and human adrenocorticotropin (ACTH). The results showed that many CRH-like immunoreactive neurons were distributed specifically on the dorsal side and ventral side of brain vesicle, while a few CRH-like neurons and their fibers in spinal cord. At the same time, the epithelial cells in the basic region of Hatschek’s pit were shown immunopositive to CRH antibody. In gonads (ovary and testis), CRH-immunopositive substance was localized in the cytoplasm near oocyte nucleus and in early spermatogenic cells. ACTH-like immunoreactivities were observed specially in the neurons and their protrusions localized on the ventral side of the brain vesicle and in spinal cord, and also in epithelial cells of Hatschek’s pit, enteric neurons of digestive tract, oocytes in ovary and in early spermatogenic cells as well. It was found for the first time that CRH-like neurons existed in the middle region of brain vesicle (corresponding to the hypothalamus of vertebrates) and ACTH-like immunopositive cells existed in Hatschek’s pit, implying that a control mechanism between brain vesicle and Hatschek’s pit maybe had been already built in amphioxus as that in vertebrates. The present study will provide new morphological evidence for the origin and evolution of ACTH. In addition, the immunoreactivities of CRH and ACTH in the digestive tract and gonads suggested other physiological function of CRH and ACTH in amphioxus.     

mmunohistochemical localization of gonadotropin-releasing hormone receptors (GnRHR) in the nervous system, Hatschek's pit and gonads of amphioxus, Branchiostoma belcheri

Using gonadotropin-releasing hormone (GnRH) anti-idiotypic antibodies and APA im-munohistochemical method, the immunoreactivity of GnRHR in the nervous system, Hatschek's pit and gonads of amphioxus has been located. It is found for the first time that the immunoreactivity of GnRHR exists in the nerve cells and fibers in the amphioxus's brain and nerve tube and the epithelial cells of Hatschek's pit at the different stages of gonadal development. At the same time, it is also found that GnRHR also exists in the ovary and testis of different developed stages. These findings provide morphological new proof for the informative transfer and regulation between brain and Hatschek's pit mediation by GnRHR, and for the understanding of the mechanism of action on the reproductive endocrine control axis among brain-Hatschek's pit-go-nads.     

Localization and expression of LHβmRNA in the brain and Hatschek's pit of amphioxus, Branchiostoma belcheri

To determine whether gonadotropin-like substance in the brain and Hatschek' s pit of amphioxus issynthesized by the tissue in situ or transported from other tissue, a histochemical study was carried out by in situ hy- bridization using digoxigenin (DIG)-labelled LHβRNAprobes. The results showed that LHβmRNA expressed in the nerve cells of brain and the epithelial cells of Hatschek' s pit, thus providing new evidence for the homology of pituitary of ver- tebrates with Hatschek' s pit and the functional evolution of gonadotropin.     

Synergistic actions of complement and lysozyme in clearance of Escherichia coli from amphioxus Branchiostoma belcheri

Amphioxus appears lacking free circulating blood cells. How it clears invading pathogens from its body remains unknown to date. We demonstrate here that amphioxus Branchiostoma belcheri is capable of efficiently eliminating the invading bacterium Escherichia coli from its humoral fluid, and the complement and lysozyme are both involved in the elimination of the invading pathogen. Both the complement and lysozyme act in concert against the invading bacterium, but the complement appears playing a more dominant role than the lysozyme.