PMID- 32966893
OWN - NLM
STAT- MEDLINE
DCOM- 20210505
LR  - 20210505
IS  - 1095-9947 (Electronic)
IS  - 1050-4648 (Linking)
VI  - 107
IP  - Pt A
DP  - 2020 Dec
TI  - Both quiescent and proliferating cells circulate in the blood of the invasive 
      apple snail Pomacea canaliculata.
PG  - 95-103
LID - S1050-4648(20)30649-5 [pii]
LID - 10.1016/j.fsi.2020.09.026 [doi]
AB  - Gastropod hematopoiesis occurs at specialized tissues in some species, but the 
      evidence also suggests that hemocyte generation is maybe widespread in the 
      connective tissues or the blood system in others. In Ampullariidae 
      (Caenogastropoda), both the kidney and the lung contain putative hematopoietic 
      cells, which react to immune challenges. In the current study, we wanted to 
      explore if hematopoiesis occurs in the blood of Pomacea canaliculata. Thus, we 
      obtained circulating hemocytes from donor animals and tested their ability to 
      proliferate in the blood of conspecific recipients. We tracked cell proliferation 
      by labeling the donors' hemocytes with the fluorescent cell proliferation marker 
      carboxyfluorescein diacetate succinimidyl ester (CFSE). Transferred CFSE-labeled 
      hemocytes survived and proliferated into the recipients' circulation for at least 
      17 days. We also determined the cell cycle status of circulating hemocytes by 
      using the propidium iodide (PI) and acridine orange (AO) staining methods. Flow 
      cytometry analyses showed that most PI-stained hemocytes were in the G1 phase 
      (~96%), while a lower proportion of cells were through the G2/S-M transition 
      (~4%). When we instead used AO-staining, we further distinguished a subpopulation 
      of cells (~5%) of low size, complexity-granularity, and RNA content. We regarded 
      this subpopulation as quiescent cells. In separate experimental sets, we 
      complemented these findings by assessing in circulating hemocytes two 
      evolutionary conserved features of quiescent, undifferentiated cells. First, we 
      used JC-1 staining to determine the mitochondrial membrane potential (Psi(m)) of 
      circulating hemocytes, which is expected to be low in quiescent cells. Most 
      hemocytes (~87%) showed high aggregation of JC-1, which indicates a high Psi(m). 
      Besides that, a small hemocyte subpopulation (~11%) showed low aggregation of the 
      dye, thus indicating a low Psi(m). It is known that the transition from a quiescent 
      to a proliferating state associates with an increase of the Psi(m). The specificity 
      of these changes was here controlled by membrane depolarization with the Psi(m) 
      disruptor CCCP. Second, we stained hemocytes with Hoechst33342 dye to determine 
      the efflux activity of ABC transporters, which participate in the multixenobiotic 
      resistance system characteristic of undifferentiated cells. Most hemocytes (>99%) 
      showed a low dye-efflux activity, but a small proportion of cells (0.06-0.12%) 
      showed a high dye-efflux activity, which was significantly inhibited by 100 and 
      500 muM verapamil, and thus is indicative of an undifferentiated subpopulation of 
      circulating hemocytes. Taken together, our results suggest that, among 
      circulating hemocytes, there are cells with the ability to proliferate or to stay 
      in a quiescent state and behave as progenitor cells later, either in the 
      circulation or the hematopoietic tissues/organs.
CI  - Copyright (c) 2020 Elsevier Ltd. All rights reserved.
FAU - Rodriguez, Cristian
AU  - Rodriguez C
AD  - IHEM, CONICET, Universidad Nacional de Cuyo, Mendoza, Argentina; Universidad 
      Nacional de Cuyo, Facultad de Ciencias Medicas, Instituto de Fisiologia, Mendoza, 
      Argentina; Universidad Nacional de Cuyo, Facultad de Ciencias Exactas y 
      Naturales, Departamento de Biologia, Mendoza, Argentina.
FAU - Simon, Valeska
AU  - Simon V
AD  - Centro de Medicina Regenerativa, Facultad de Medicina Clinica Alemana Universidad 
      del Desarrollo, 7710162, Santiago, Chile.
FAU - Conget, Paulette
AU  - Conget P
AD  - Centro de Medicina Regenerativa, Facultad de Medicina Clinica Alemana Universidad 
      del Desarrollo, 7710162, Santiago, Chile. Electronic address: pconget@udd.cl.
FAU - Vega, Israel A
AU  - Vega IA
AD  - IHEM, CONICET, Universidad Nacional de Cuyo, Mendoza, Argentina; Universidad 
      Nacional de Cuyo, Facultad de Ciencias Medicas, Instituto de Fisiologia, Mendoza, 
      Argentina; Universidad Nacional de Cuyo, Facultad de Ciencias Exactas y 
      Naturales, Departamento de Biologia, Mendoza, Argentina. Electronic address: 
      ivega@mendoza-conicet.gob.ar.
LA  - eng
PT  - Journal Article
DEP - 20200920
PL  - England
TA  - Fish Shellfish Immunol
JT  - Fish & shellfish immunology
JID - 9505220
SB  - IM
MH  - Animals
MH  - Cell Count
MH  - Flow Cytometry
MH  - Hematopoiesis/*immunology
MH  - Hemocytes/*immunology
MH  - Introduced Species
MH  - Snails/*immunology
OTO - NOTNLM
OT  - Angiostrongylus cantonensis
OT  - Cell cycle
OT  - Hematopoiesis
OT  - Hemocyte transfer
OT  - Invasive species
OT  - Mitochondrial membrane potential
OT  - Multixenobiotic resistance system
OT  - Stem cell
EDAT- 2020/09/24 06:00
MHDA- 2021/05/06 06:00
CRDT- 2020/09/23 20:05
PHST- 2020/06/16 00:00 [received]
PHST- 2020/09/13 00:00 [revised]
PHST- 2020/09/17 00:00 [accepted]
PHST- 2020/09/24 06:00 [pubmed]
PHST- 2021/05/06 06:00 [medline]
PHST- 2020/09/23 20:05 [entrez]
AID - S1050-4648(20)30649-5 [pii]
AID - 10.1016/j.fsi.2020.09.026 [doi]
PST - ppublish
SO  - Fish Shellfish Immunol. 2020 Dec;107(Pt A):95-103. doi: 
      10.1016/j.fsi.2020.09.026. Epub 2020 Sep 20.