Functional differentiation of spider hemocytes by light and transmission electron microscopy, and MALDI-MS-imaging

Kuhn‐Nentwig, Lucia; Kopp, Lukas S.; Nentwig, Wolfgang; Haenni, Beat; Streitberger, Kathrin; Schürch, Stefan; Schaller, Johann

doi:10.1016/j.dci.2013.10.008

Cited by 25 publications

(15 citation statements)

References 30 publications

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“…Anticoagulants commonly used for hematology in domestic species such as lithium heparin, sodium citrate, and EDTA are reportedly inefficient in preventing coagulation in invertebrates, consistent with our findings when lithium heparin was used in the present study. Various solutions have been reported effective in preventing the coagulation of arachnid hemolymph, but these solutions are not readily available for clinicians. Therefore, the simpler technique of rapid hemolymph cooling was investigated, which sufficiently slowed coagulation to permit immediate analysis.…”

Section: Discussionmentioning

confidence: 99%

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Hemolymph cytology, hemocyte count, glucose, and electrolyte reference intervals in 93 Cameroon red tarantulas (Hysterocrates gigas)

Larouche

Beeler-Marfisi

Attard

et al. 2019

Veterinary Clinical Pathol

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Background: Theraphosidae is a large family of tarantula spiders commonly kept as pets or display animals by zoological institutions. Interest and demand for arachnid medicine have grown over the last decade. While hemolymph analysis could be a fundamental tool for arachnid health assessment, RIs are generally lacking for arachnid species.Objectives: The objectives of this study were to describe the appearance of hemocytes and establish RIs for total and differential hemocyte counts as well as hemolymph glucose and electrolyte concentrations in the Cameroon red tarantula (CRT) (Hysterocrates gigas).Methods: Ninety-three CRTs were anesthetized using isoflurane or sevoflurane, and hemolymph was collected from the heart. Hemocyte counts were performed using a hemocytometer and trypan blue stain, and differential cell counts were estimated using light microscopy and a Wright stain. Hemocytes were also characterized with transmission electron microscopy (TEM). Hemolymph glucose and electrolyte concentrations were obtained using a blood gas analyzer. The American Society for Veterinary Clinical Pathology (ASVCP) guidelines were used to calculate RIs. Results:The most abundant hemocytes were plasmatocytes, followed by granulocytes, cyanocytes, and prohemocytes. Hemocytes were characterized by TEM except for cyanocytes. All RIs had a broad range, likely due to a combination of population heterogeneity, high biologic variability of the species, and the propensity of samples to clot. Conclusions:The current study is the first to establish RIs for hemolymph cytologic parameters with a sample size of over 20 individuals in any theraphosid species. RIs for CRT hemolymph biochemical analytes are also reported. K E Y W O R D S arachnids, electrolytes, hemocyte, hemolymph, reference intervals, tarantulas 1 | INTRODUC TI ON Theraphosid tarantulas are commonly kept as pets by hobbyists and as display animals by zoological institutions. The Cameroon red tarantula (CRT) (Hysterocrates gigas), also known as the giant baboon spider, is a burrowing species that originates from Cameroon in Western Africa. Compared with females, adult males have a smaller opisthosoma, and the ends of their pedipalp tarsi

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Section: Discussionmentioning

confidence: 99%

“…A few studies have evaluated the biochemical composition and osmolality of the hemolymph of theraphosid spiders, and other studies have described the morphology and function of hemocytes in various spider species . However, publications reporting reference values for total and differential hemocyte counts are lacking.…”

Section: Introductionmentioning

confidence: 99%

Hemolymph cytology, hemocyte count, glucose, and electrolyte reference intervals in 93 Cameroon red tarantulas (Hysterocrates gigas)

Larouche

Beeler-Marfisi

Attard

et al. 2019

Veterinary Clinical Pathol

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“…Moreover, a 30 kDa protein from the hemolymph of B. mori (silkworm) could protect Sf-9 cells from apoptosis, inhibiting the activation triggered by Sf-Caspase-I, and increasing cell viability in the culture (Rhee et al, 2002). Recently, two studies characterized C. salei hemocytes and the hematopoiesis and hematopoietic organs in arthropods (Kuhn-Nentwig et al, 2014;Grigorian and Hartenstein, 2013;respectively). As discussed by Kuhn-Nentwig, main aspects of the immune system in arachnids, such as the number and type of hemocyte differ astonishingly among the various arachnid groups, making extrapolations and prognoses very difficult.…”

Section: Q3mentioning

confidence: 96%

Characterization of Brown spider (Loxosceles intermedia) hemolymph: Cellular and biochemical analyses

Bednaski

Trevisan-Silva

Matsubara

et al. 2015

Toxicon

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This is the first study on the hemolymph from a spider of the Loxosceles genus. These animals are responsible for a great number of envenomation cases worldwide. Several studies on Loxosceles venoms have been published, and the knowledge about the venom and its toxins is considerable, not only regarding the biological and biochemical characterization, but also regarding structural, genetic and phylogenetic approaches. However, the literature on Loxosceles hemolymph is nonexistent. The main goal of the present study was to characterize biochemically the hemolymph content, and especially, to identify its different hemocytes. Moreover, many papers have already shown molecules whose source is the hemolymph and their very interesting activities and biomedical applications, for example, antifungal and antibacterial activities. A 2D-SDS-PAGE of brown spider hemolymph showed approximately 111 spots for pH 3-10 and 150 spots for pH 4-7. A lectin-blotting assay showed that hemolymph carbohydrate residues were similar to those found in venom. Several types of TAG and DAG phospholipids were found in the hemolymph and characterized by HPTLC and mass spectrometry. Four different hemocytes were characterized in Loxosceles intermedia hemolymph: prohemocyte, plasmatocyte, granulocyte and adipohemocyte. This paper opens new possibilities on toxinology, studying an unknown biological material, and it characterizes a source of molecules with putative biotechnological applications.

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“…Future research can benefit from MALDI imaging mass spectrometry (78, 79) to investigate E3-mediated production of HDPs in vivo . Combining MALDI imaging with the recently developed ubiquitin variants (80), which systematically modulate HECT E3 ligase activity, as well as proteasome inhibitors (81), will be useful to probe the mechanism of HDP production proposed herein.…”

Section: Discussionmentioning

confidence: 99%

Evidence of an Antimicrobial Peptide Signature Encrypted in HECT E3 Ubiquitin Ligases

et al. 2017

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The ubiquitin-proteasome pathway (UPP) is a hallmark of the eukaryotic cell. In jawed vertebrates, it has been co-opted by the adaptive immune system, where proteasomal degradation produces endogenous peptides for major histocompatibility complex class I antigen presentation. However, proteolytic products are also necessary for the phylogenetically widespread innate immune system, as they often play a role as host defense peptides (HDPs), pivotal effectors against pathogens. Here, we report the identification of the arachnid HDP oligoventin, which shares homology to a core member of the UPP, E3 ubiquitin ligases. Oligoventin has broad antimicrobial activity and shows strong synergy with lysozymes. Using computational and phylogenetic approaches, we show high conservation of the oligoventin signature in HECT E3s. In silico simulation of HECT E3s self-proteolysis provides evidence that HDPs can be generated by fine-tuned 26S proteasomal degradation, and therefore are consistent with the hypothesis that oligoventin is a cryptic peptide released by the proteolytic processing of an Nedd4 E3 precursor protein. Finally, we compare the production of HDPs and endogenous antigens from orthologous HECT E3s by proteasomal degradation as a means of analyzing the UPP coupling to metazoan immunity. Our results highlight the functional plasticity of the UPP in innate and adaptive immune systems as a possibly recurrent mechanism to generate functionally diverse peptides.

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Functional differentiation of spider hemocytes by light and transmission electron microscopy, and MALDI-MS-imaging

Cited by 25 publications

References 30 publications

Hemolymph cytology, hemocyte count, glucose, and electrolyte reference intervals in 93 Cameroon red tarantulas (Hysterocrates gigas)

Hemolymph cytology, hemocyte count, glucose, and electrolyte reference intervals in 93 Cameroon red tarantulas (Hysterocrates gigas)

Characterization of Brown spider (Loxosceles intermedia) hemolymph: Cellular and biochemical analyses

Evidence of an Antimicrobial Peptide Signature Encrypted in HECT E3 Ubiquitin Ligases

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