Immunohistochemical study of subepidermal connective of molluscan integument

Sections of integument from gastropod, bivalve and cephalopod species were studied immunohistochemically to determine reactivity to antibody against the type I-like collagen from Sepia cartilage and antibodies against components of the extracellular matrix (ECM) of vertebrate connective tissue: type I, III, IV, V, and VI collagens, laminin, nidogen and heparan sulphate. All samples exhibited similar reactivities to the antibodies, although differences in the intensity and localization of the immunostaining were found that were clearly correlated with between-species differences in integumental ultrastructure. These findings indicate that the composition of the integumental ECM is similar in the three classes of molluscs examined and that several types of collagen are present. However molluscan subepidermal connective tissue differs from the ECM of vertebrate dermis: molluscan integumental ECM contains collagens similar to type I, V and VI collagens but has no type III-similar collagen. Furthermore molecules similar to the type IV collagen, laminin, nidogen and heparan sulphate of vertebrates were present ubiquitously in molluscan basement membrane, confirming the statement that the structure and composition of basement membrane have remained constant throughout the evolution of all animal phyla

The cytoskeleton of chondrocytes of Sepia officinalis (Mollusca, Cephalopoda): an immunocytochemical study.

Our previous electron microscope study showed that chondrocytes from cephalopod cartilage possess a highly developed cytoskeleton and numerous cytoplasmic processes that ramify extensively through the tissue. We have now carried out a light microscope immunocytochemical study of chondrocytes from the orbital cartilage of Sepia officinalis to obtain indications as to the nature of the cytoskeletal components. We found clear positivity to antibodies against mammalian tubulin, vimentin, GFAP, and actin, but not keratin. The simultaneous presence of several cytoskeletal components is consistent with the hypothesis that cephalopod chondrocytes have the characteristics of both chondrocytes and osteocytes of vertebrates, which endow the tissue as a whole with some of the properties of vertebrate bone. We confirm, therefore, the presence in molluscs of the ubiquitous cytoskeletal proteins of metazoan cells that have remained highly conserved throughout phylogenetic evolution

Lycopersicon esculentum lectin: an effective and versatile endothelial marker of normal and tumoral blood vessels in the central nervous system

The binding of Lycopersicon esculentum lectin (LEA) to the vascular endothelium was studied in the central nervous system of rat, mouse and guinea pig at different developmental ages, and in a gliosarcoma model. Our observations showed that LEA consistently stained the entire vascular tree in the spinal cord and in the brain of all animal species at all developmental ages investigated. In the tumor model, the staining of the vascular network was very reproducible, enabled an easy identification of vascular profiles and displayed a higher efficiency when compared to two other commonly used vascular marker (EHS laminin and PECAM-1). Moreover, our results showed that LEA staining was comparable in both vibratome and paraffin sections and could be easily combined with other markers in double labeling experiments. These observations indicate that LEA staining may represent an effective and versatile endothelial marker for the study of the vasculature of the central nervous system in different animal species and experimental conditions

Erythropoietin Amplifies Stroke-Induced Oligodendrogenesis in the Rat

Erythropoietin (EPO), a hematopoietic cytokine, enhances neurogenesis and angiogenesis during stroke recovery. In the present study, we examined the effect of EPO on oligodendrogenesis in a rat model of embolic focal cerebral ischemia.Recombinant human EPO (rhEPO) at a dose of 5,000 U/kg (n = 18) or saline (n = 18) was intraperitoneally administered daily for 7 days starting 24 h after stroke onset. Treatment with rhEPO augmented actively proliferating oligodendrocyte progenitor cells (OPCs) measured by NG2 immunoreactive cells within the peri-infarct white matter and the subventricular zone (SVZ), but did not protect against loss of myelinating oligodendrocytes measured by cyclic nucleotide phosphodiesterase (CNPase) positive cells 7 days after stroke. However, 28 and 42 days after stroke, treatment with rhEPO significantly increased myelinating oligodendrocytes and myelinated axons within the peri-infarct white matter. Using lentivirus to label subventricular zone (SVZ) neural progenitor cells, we found that in addition to the OPCs generated in the peri-infarct white matter, SVZ neural progenitor cells contributed to rhEPO-increased OPCs in the peri-infarct area. Using bromodeoxyuridine (BrdU) for birth-dating cells, we demonstrated that myelinating oligodendrocytes observed 28 days after stroke were derived from OPCs. Furthermore, rhEPO significantly improved neurological outcome 6 weeks after stroke. In vitro, rhEPO increased differentiation of adult SVZ neural progenitor cells into oligodendrocytes and enhanced immature oligodendrocyte cell proliferation.Our in vivo and in vitro data indicate that EPO amplifies stroke-induced oligodendrogenesis that could facilitate axonal re-myelination and lead to functional recovery after stroke

Hydrolyzed eggshell membrane immobilized on phosphorylcholine polymer supplies extracellular matrix environment for human dermal fibroblasts

We have found that a water-soluble alkaline-digested form of eggshell membrane (ASESM) can provide an extracellular matrix (ECM) environment for human dermal fibroblast cells (HDF) in vitro. Avian eggshell membrane (ESM) has a fibrous-meshwork structure and has long been utilized as a Chinese medicine for recovery from burn injuries and wounds in Asian countries. Therefore, ESM is expected to provide an excellent natural material for biomedical use. However, such applications have been hampered by the insolubility of ESM proteins. We have used a recently developed artificial cell membrane biointerface, 2-methacryloyloxyethyl phosphorylcholine polymer (PMBN) to immobilize ASESM proteins. The surface shows a fibrous structure under the atomic force microscope, and adhesion of HDF to ASESM is ASESM-dose-dependent. Quantitative mRNA analysis has revealed that the expression of type III collagen, matrix metalloproteinase-2, and decorin mRNAs is more than two-fold higher when HDF come into contact with a lower dose ASESM proteins immobilized on PMBN surface. A particle-exclusion assay with fixed erythrocytes has visualized secreted water-binding molecules around the cells. Thus, HDF seems to possess an ECM environment on the newly designed PMBN-ASESM surface, and future applications of the ASESM-PMBN system for biomedical use should be of great interest

The cytoskeleton of chondrocytes of Sepia officinalis (Mollusca, Cephalopoda): an immunocytochemical study.

Our previous electron microscope study showed that chondrocytes from cephalopod cartilage possess a highly developed cytoskeleton and numerous cytoplasmic processes that ramify extensively through the tissue. We have now carried out a light microscope immunocytochemical study of chondrocytes from the orbital cartilage of Sepia officinalis to obtain indications as to the nature of the cytoskeletal components. We found clear positivity to antibodies against mammalian tubulin, vimentin, GFAP, and actin, but not keratin. The simultaneous presence of several cytoskeletal components is consistent with the hypothesis that cephalopod chondrocytes have the characteristics of both chondrocytes and osteocytes of vertebrates, which endow the tissue as a whole with some of the properties of vertebrate bone. We confirm, therefore, the presence in molluscs of the ubiquitous cytoskeletal proteins of metazoan cells that have remained highly conserved throughout phylogenetic evolution

Immunohistochemical study of subepidermal connective of molluscan integument

Sections of integument from gastropod, bivalve and cephalopod species were studied immunohistochemically to determine reactivity to antibody against the type I-like collagen from Sepia cartilage and antibodies against components of the extracellular matrix (ECM) of vertebrate connective tissue: type I, III, IV, V, and VI collagens, laminin, nidogen and heparan sulphate. All samples exhibited similar reactivities to the antibodies, although differences in the intensity and localization of the immunostaining were found that were clearly correlated with between-species differences in integumental ultrastructure. These findings indicate that the composition of the integumental ECM is similar in the three classes of molluscs examined and that several types of collagen are present. However molluscan subepidermal connective tissue differs from the ECM of vertebrate dermis: molluscan integumental ECM contains collagens similar to type I, V and VI collagens but has no type III-similar collagen. Furthermore molecules similar to the type IV collagen, laminin, nidogen and heparan sulphate of vertebrates were present ubiquitously in molluscan basement membrane, confirming the statement that the structure and composition of basement membrane have remained constant throughout the evolution of all animal phyla

Immunohistochemical study of subepidermal connective of molluscan integument

Sections of integument from gastropod, bivalve and cephalopod species were studied immunohistochemically to determine reactivity to antibody against the type I-like collagen from Sepia cartilage and antibodies against components of the extracellular matrix (ECM) of vertebrate connective tissue: type I, III, IV, V, and VI collagens, laminin, nidogen and heparan sulphate. All samples exhibited similar reactivities to the antibodies, although differences in the intensity and localization of the immunostaining were found that were clearly correlated with between-species differences in integumental ultrastructure. These findings indicate that the composition of the integumental ECM is similar in the three classes of molluscs examined and that several types of collagen are present. However molluscan subepidermal connective tissue differs from the ECM of vertebrate dermis: molluscan integumental ECM contains collagens similar to type I, V and VI collagens but has no type III-similar collagen. Furthermore molecules similar to the type IV collagen, laminin, nidogen and heparan sulphate of vertebrates were present ubiquitously in molluscan basement membrane, confirming the statement that the structure and composition of basement membrane have remained constant throughout the evolution of all animal phyla