RESUMO
The harderian gland (HG) is a gland located at the base of the nictating membrane and fills the inferomedial aspect of the orbit in rodents. It is under the influence of the hypothalamic-pituitary-gonadal axis and, because of its hormone receptors, it is a target tissue for prolactin (PRL) and sex steroid hormones (estrogen and progesterone). In humans and murine, the anterior surface of the eyes is protected by a tear film synthesized by glands associated with the eye. In order to understand the endocrine changes caused by hyperprolactinemia in the glands responsible for the formation of the tear film, we used an animal model with metoclopramide-induced hyperprolactinemia (HPRL). Given the evidences that HPRL can lead to a process of cell death and tissue fibrosis, the protein expression of small leucine-rich proteoglycans (SLRPs) was analyzed through immunohistochemistry in the HG of the non- and the pregnant female mice with hyperprolactinemia. The SRLPs are related to collagen fibrillogenesis and they participate in pro-apoptotic signals. Our data revealed that high prolactin levels and changes in steroid hormones (estrogen and progesterone) can lead to an alteration in the amount of collagen, and in the structure of type I and III collagen fibers through changes in the amounts of lumican and decorin, which are responsible for collagen fibrillogenesis. This fact can lead to the impaired functioning of the HG by excessive apoptosis in the HG of the non- and the pregnant female mice with HPRL and especially in the HG of pregnancy-associated hyperprolactinemia.
Assuntos
Glândula de Harder , Hiperprolactinemia , Gravidez , Humanos , Camundongos , Feminino , Animais , Proteoglicanas/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Hiperprolactinemia/induzido quimicamente , Hiperprolactinemia/metabolismo , Proteoglicanas de Sulfatos de Condroitina/metabolismo , Decorina/metabolismo , Prolactina/efeitos adversos , Prolactina/análise , Prolactina/metabolismo , Progesterona , Glândula de Harder/metabolismo , Colágeno/metabolismo , Matriz Extracelular/metabolismo , Estrogênios/efeitos adversos , Estrogênios/análise , Estrogênios/metabolismoRESUMO
OBJECTIVE: To assess the impact of the metoclopramide-induced hyperprolactinemia in cellular death and proliferation in the harderian gland of female mice. METHODS: Twenty female mice were divided into two groups of 10 animals each and treated: 0.2 mL of saline solution (controls, Ctr) and 200 µg of metoclopramide (experimental, hyperprolactinemia), both for 50 consecutive days and at 12:00 a.m. On the 50th day, the female were euthanized, and the harderian glands were removed and processed for immunohistochemistry for detected ki67 and TUNEL method. Data were statistically analyzed by unpaired Student's t test (p<0.05). RESULTS: The harderian gland of the hyperprolactinemia group showed increase in the immunoexpression of Ki67 and TUNEL compared to the Ctr group (p<0.05), and there was no significant difference in the amount of porphyrin in the HPrl group compared to the Ctr group. CONCLUSION: The hyperprolactinemia led to increased cell death in the acini the harderian gland and cell proliferation in the stroma glandular, fact that suggesting a reduction process of cellular activity and fibrosis, which suggests impairment in the functioning of the lacrimal harderian.
Assuntos
Glândula de Harder , Hiperprolactinemia , Aparelho Lacrimal , Porfirinas , Animais , Proliferação de Células , Feminino , Hiperprolactinemia/induzido quimicamente , CamundongosRESUMO
The ATP-Binding Cassette, subfamily G, member 2 (ABCG2) transporter is associated with the regulation of protoporphyrin IX transport and of other intermediates in heme biosynthesis. Because the hamster Harderian gland (HG) exhibits high concentrations of porphyrins and sexual dimorphism, we analyzed the hamster ABCG2. Cloned cDNA [2098-base pairs (bp)] contains an open-reading frame (ORF) of 1971-bp that encodes a 656 amino-acid protein with a molecular weight of 72844.56Da. The hamster ABCG2 sequence is conserved phylogenetically and shares a high percentage of identity with mouse (89%), rat (88%), and human (84%) transporters. Within its structure, a Walker A (G-X-X-G-X-G-K-S), a C signature motif characteristic of ABC transporters, and six putative transmembrane domains (TMDs) were identified. ABCG2 mRNA was detected in all hamster tissues, with higher amounts found in HG, brain, cerebellum, kidney, gut, ovary, and testis. Harderian ABCG2 expression exhibits a sexually dimorphic pattern where females display higher mRNA levels than males. Different patterns of transcriptional profiles of ABCG2 during the estrous cycle and after gonadectomy in both sexes were also observed. The differential expression between male and female HGs suggests that ABCG2 is under the regulation of gonadal steroids. The ABCG2 transporter is likely involved in the endogenous regulation of porphyrins in hamster HGs.
Assuntos
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Hormônios Esteroides Gonadais/metabolismo , Glândula de Harder/metabolismo , Protoporfirinas/metabolismo , Animais , Cricetinae , DNA Complementar , Feminino , Humanos , Masculino , Mesocricetus , Camundongos , Porfirinas/metabolismo , RNA Mensageiro , Ratos , Caracteres SexuaisRESUMO
The Harderian gland is a cephalic structure, widely distributed among vertebrates. In snakes, the Harderian gland is anatomically connected to the vomeronasal organ via the nasolacrimal duct, and in some species can be larger than the eyes. The function of the Harderian gland remains elusive, but it has been proposed to play a role in the production of saliva, pheromones, thermoregulatory lipids and growth factors, among others. Here, we have profiled the transcriptomes of the Harderian glands of three non-front-fanged colubroid snakes from Cuba: Caraiba andreae (Cuban Lesser Racer); Cubophis cantherigerus (Cuban Racer); and Tretanorhinus variabilis (Caribbean Water Snake), using Illumina HiSeq2000 100â¯bp paired-end. In addition to ribosomal and non-characterized proteins, the most abundant transcripts encode putative transport/binding, lipocalin/lipocalin-like, and bactericidal/permeability-increasing-like proteins. Transcripts coding for putative canonical toxins described in venomous snakes were also identified. This transcriptional profile suggests a more complex function than previously recognized for this enigmatic organ.
Assuntos
Colubridae/metabolismo , Regulação da Expressão Gênica/fisiologia , Glândula de Harder/metabolismo , Proteínas de Répteis/biossíntese , Venenos de Serpentes/biossíntese , Transcriptoma/fisiologia , Animais , Colubridae/genética , Cuba , Proteínas de Répteis/genética , Venenos de Serpentes/genéticaRESUMO
The objective of this study was to determine how cytokine transcription profiles correlate with patterns of infectious laryngotracheitis virus (ILTV) replication in the trachea, Harderian gland, and trigeminal ganglia during the early and late stages of infection after intratracheal inoculation. Viral genomes and transcripts were detected in the trachea and Harderian gland but not in trigeminal ganglia. The onset of viral replication in the trachea was detected at day one post-infection and peaked by day three post-infection. The peak of pro-inflammatory (CXCLi2, IL-1ß, IFN-γ) and anti-inflammatory (IL-13, IL-10) cytokine gene transcription, 5 days post-infection, coincided with the increased recruitment of inflammatory cells, extensive tissue damage, and limiting of virus replication in the trachea. In contrast, transcription of the IFN-ß gene in the trachea remained unaffected suggesting that ILTV infection blocks type I interferon responses. In the Harderian gland, the most evident transcription change was the early and transient upregulation of the IFN-γ gene at 1 day post-infection, which suggests that the Harderian gland is prepared to rapidly respond to ILTV infection. Overall, results from this study suggest that regulation of Th1 effector cells and macrophage activity by Th1/2 cytokines was pertinent to maintain a balanced immune response capable of providing an adequate Th1-mediated protective immunity, while sustaining some immune homeostasis in preparation for the regeneration of the tracheal mucosa.
Assuntos
Citocinas/metabolismo , Glândula de Harder/metabolismo , Infecções por Herpesviridae/veterinária , Herpesvirus Galináceo 1/patogenicidade , Traqueia/metabolismo , Gânglio Trigeminal/metabolismo , Animais , Galinhas , Citocinas/genética , DNA , Regulação da Expressão Gênica/imunologia , Genoma Viral , Glândula de Harder/virologia , Infecções por Herpesviridae/metabolismo , Infecções por Herpesviridae/virologia , Herpesvirus Galináceo 1/fisiologia , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/metabolismo , Doenças das Aves Domésticas/virologia , RNA , Organismos Livres de Patógenos Específicos , Traqueia/virologia , Transcrição Gênica , Gânglio Trigeminal/virologia , Carga Viral , Virulência , Replicação ViralRESUMO
Protoporphyrin IX (PpIX) is a precursor of heme synthesis and is known to be an active photosensitizer and precursor of photosensitizers applied in photodynamic therapy (PDT) and photodynamic diagnostics (PDD). On irradiation with visible light, PpIX undergoes phototransformation, producing photoproducts which may also be phototoxic and increase its efficacy. The mechanism of PpIX phototransformation depends on environmental characteristics and can be different in vitro and in vivo. In this paper, we present a comparative study of the photoactivity of synthetic PpIX and PpIX extracted from the Harderian gland of ssp Rattus novergicus albinus rats, along with their photoproducts toward murine B16F-10 melanoma cells. It was observed that when irradiated with visible light the endogenous PpIX demonstrates photocytotoxicity ten times higher than the synthetic PpIX. The photoproduct of endogenous PpIX also possesses phototoxicity, though slightly lower than that of PpIX itself. The rate of cell internalization for both endogenous PpIX and its photoproduct was eightfold greater than that obtained for the synthetic porphyrin. This difference might result from a complexation of the native PpIX with some amphiphilic compounds during its synthesis within the Harderian glands, which facilitates the cell uptake of PpIX. Fluorescence microscopy images show that both endogenous and synthetic porphyrins are localized after uptake predominantly in the mitochondrial region of cells.
Assuntos
Melanoma/patologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Protoporfirinas/farmacologia , Animais , Transporte Biológico , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Escuridão , Glândula de Harder/metabolismo , Espaço Intracelular/metabolismo , Masculino , Melanoma/tratamento farmacológico , Camundongos , Fármacos Fotossensibilizantes/síntese química , Fármacos Fotossensibilizantes/isolamento & purificação , Fármacos Fotossensibilizantes/metabolismo , Protoporfirinas/síntese química , Protoporfirinas/isolamento & purificação , Protoporfirinas/metabolismo , RatosRESUMO
The aim of the present study was to demonstrate the histological and histochemical structure of the Harderian gland in wild and hybrid of wild and domestic birds. The samples were stained with haematoxylin-eosin, methyl green-pyronin Y, periodic acid-Schiff, alcian blue pH 2.5, aldehyde fuchsin and Hale's dialyzed iron staining's. In both species, the glands had multilobar tubuloacinar structure type I. The Harderian gland was located in the orbit near the inter-orbital septum, between the medial rectus muscle, the pyramidal muscle of the third eyelid, and the ventral oblique muscle. In the common pheasant, the gland was wider in the proximal and distal part. The common pheasant had more elongated lobes of the Harderian gland than in the hybrid. In the common pheasant, the glandular cells presented darkly-stained serous secretion and lightly-stained mucous secretion. In the hybrid, the glandular cells presented seromucous secretion. The central lobar space, interacinar space, and apical parts of the acini of the Harderian glands were filled with many lymphocytes and plasma cells, particularly in the common pheasant, where centers of all large lobes were abundantly filled with plasma cells. The plasma cells dominated in common pheasant's Harderian gland, while in the hybrid, lymphocytes and plasma cells were present at similar quantities. The cells positive for periodic acid of Schiff staining were dominant in hybrid. Periodic acid-Schiff, Hale's dialyzed iron and alcian blue pH 2.5 stainings demonstrated acid-carboxylated mucopolysaccharides in the glandular cells cytoplasm of the examined birds.(AU)
Assuntos
Animais , Aves/anatomia & histologia , Aves/classificação , Glândula de Harder/anatomia & histologiaRESUMO
The aim of the present study was to demonstrate the histological and histochemical structure of the Harderian gland in wild and hybrid of wild and domestic birds. The samples were stained with haematoxylin-eosin, methyl green-pyronin Y, periodic acid-Schiff, alcian blue pH 2.5, aldehyde fuchsin and Hale's dialyzed iron staining's. In both species, the glands had multilobar tubuloacinar structure type I. The Harderian gland was located in the orbit near the inter-orbital septum, between the medial rectus muscle, the pyramidal muscle of the third eyelid, and the ventral oblique muscle. In the common pheasant, the gland was wider in the proximal and distal part. The common pheasant had more elongated lobes of the Harderian gland than in the hybrid. In the common pheasant, the glandular cells presented darkly-stained serous secretion and lightly-stained mucous secretion. In the hybrid, the glandular cells presented seromucous secretion. The central lobar space, interacinar space, and apical parts of the acini of the Harderian glands were filled with many lymphocytes and plasma cells, particularly in the common pheasant, where centers of all large lobes were abundantly filled with plasma cells. The plasma cells dominated in common pheasant's Harderian gland, while in the hybrid, lymphocytes and plasma cells were present at similar quantities. The cells positive for periodic acid of Schiff staining were dominant in hybrid. Periodic acid-Schiff, Hale's dialyzed iron and alcian blue pH 2.5 stainings demonstrated acid-carboxylated mucopolysaccharides in the glandular cells cytoplasm of the examined birds.
Assuntos
Animais , Aves/anatomia & histologia , Aves/classificação , Glândula de Harder/anatomia & histologiaRESUMO
The macroscopic anatomy and the microscopic and ultrastructural features of the Harderian gland (HG), lacrimal gland (LG) and superficial gland of the third eyelid (SGTE) of the adult European bison are described. In addition, morphometric studies were conducted and were followed by statistical analysis of the results. Tissue sections were stained with hematoxylin and eosin, methyl green-pyronin Y, periodic acid-Schiff, Alcian blue pH 2.5, aldehyde fuchsin and Hale's dialysed iron. Analysis of the staining showed that the HG has a multilobular tubuloalveolar structure with mixed secretion. The LG and the SGTE have a multilobar tubuloacinar structure with serous secretion in the LG and mucoserous in the SGTE. The TEM study demonstrates that the secretory cells of the HG, LG and SGTE have similar ultrastructural appearance, with two types of secretory vesicles in the cytoplasm of all studied glands. The histochemical staining methods and the TEM study revealed the secretory activity in the HG, LG and SGTE ducts. The structural studies can be important for establishing relations between morphological structure and functions of these glands. It can have clinical implications especially when taking into consideration the protective mechanisms of the eye.(AU)
Assuntos
Animais , Bison/anatomia & histologia , Glândula de Harder/anatomia & histologia , Aparelho Lacrimal/anatomia & histologia , Pálpebras/anatomia & histologiaRESUMO
The macroscopic anatomy and the microscopic and ultrastructural features of the Harderian gland (HG), lacrimal gland (LG) and superficial gland of the third eyelid (SGTE) of the adult European bison are described. In addition, morphometric studies were conducted and were followed by statistical analysis of the results. Tissue sections were stained with hematoxylin and eosin, methyl green-pyronin Y, periodic acid-Schiff, Alcian blue pH 2.5, aldehyde fuchsin and Hale's dialysed iron. Analysis of the staining showed that the HG has a multilobular tubuloalveolar structure with mixed secretion. The LG and the SGTE have a multilobar tubuloacinar structure with serous secretion in the LG and mucoserous in the SGTE. The TEM study demonstrates that the secretory cells of the HG, LG and SGTE have similar ultrastructural appearance, with two types of secretory vesicles in the cytoplasm of all studied glands. The histochemical staining methods and the TEM study revealed the secretory activity in the HG, LG and SGTE ducts. The structural studies can be important for establishing relations between morphological structure and functions of these glands. It can have clinical implications especially when taking into consideration the protective mechanisms of the eye.