Minichromosome maintenance protein 5 (MCM5), a member of the microchromosomal maintenance protein family, plays an important role in the initiation and extension of DNA replication. However, its role in neural development in zebrafish remains unclear. Here, we used morpholino (MO) and CRISPR/Cas9 to knock down mcm5 and investigated the developmental features of facial motor neurons (FMNs) in the hindbrain of zebrafish. We found that knockdown of mcm5 using mcm5 MO resulted in a small head, small eyes, and a blurred midbrain-hindbrain boundary, while MO injection of mcm5 led to decrease in FMNs and their migration disorder. However, the mutant of mcm5 only resulted in the migration defect of FMNs rather than quantity change. We further investigated the underlying mechanism of mcm5 in the development of hindbrain using in situ hybridization (ISH) and fgfr1a mRNA co-injected with mcm5 MO. Results from ISH showed that the fibroblast growth factor (FGF) signaling pathway was changed when the MCM5 function was lost, with the decrease in fgfr1a and the increase in fgf8, while that of pea3 had opposite trend. FMN development defects were rescued by fgfr1a mRNA co-injected with mcm5 MO. Our results demonstrated that FGF signaling pathway is required for FMN development in zebrafish. Specifically, mcm5 regulates FMN development during zebrafish growing.

1.
Cubedo
N
,
Cerdan
E
,
Sapede
D
,
Rossel
M
.
Cxcr4 and cxcr7 cooperate during tangential migration of facial motoneurons
.
Mol Cell Neurosci
.
2009
;
40
:
474
84
. .
2.
Zannino
DA
,
Sagerstrom
CG
,
Appel
B
.
Olig2-expressing hindbrain cells are required for migrating facial motor neurons
.
Dev Dyn
.
2012
;
241
:
315
26
. .
3.
Cooper
KL
,
Armstrong
J
,
Moens
CB
.
Zebrafish foggy/spt 5 is required for migration of facial branchiomotor neurons but not for their survival
.
Dev Dyn
.
2005
;
234
:
651
8
. .
4.
Wiellette
EL
,
Sive
H
.
Early requirement for fgf8 function during hindbrain pattern formation in zebrafish
.
Dev Dyn
.
2004
;
229
:
393
9
. .
5.
Selland
LG
,
Koch
S
,
Laraque
M
,
Waskiewicz
AJ
.
Coordinate regulation of retinoic acid synthesis by pbx genes and fibroblast growth factor signaling by hoxb1b is required for hindbrain patterning and development
.
Mech Dev
.
2018
;
150
:
28
41
. .
6.
Coster
G
,
Frigola
J
,
Beuron
F
,
Morris
EP
,
Diffley
JF
.
Origin licensing requires ATP binding and hydrolysis by the mcm replicative helicase
.
Mol Cell
.
2014
;
55
:
666
77
. .
7.
Edwards
MC
,
Tutter
AV
,
Cvetic
C
,
Gilbert
CH
,
Prokhorova
TA
,
Walter
JC
.
Mcm2-7 complexes bind chromatin in a distributed pattern surrounding the origin recognition complex in xenopus egg extracts
.
J Biol Chem
.
2002
;
277
:
33049
57
. .
8.
Ryu
S
,
Driever
W
.
Minichromosome maintenance proteins as markers for proliferation zones during embryogenesis
.
Cell Cycle
.
2006
;
5
(
11
):
1140
2
. .
9.
Roy
NM
,
Sagerström
CG
.
An early FGF signal required for gene expression in the zebrafish hindbrain primordium
.
Brain Res Dev Brain Res
.
2004
;
148
:
27
42
. .
10.
Sunmonu
NA
,
Li
K
,
Guo
Q
,
Li
JY
.
Gbx2 and fgf8 are sequentially required for formation of the midbrain-hindbrain compartment boundary
.
Development
.
2011
;
138
:
725
34
. .
11.
Labalette
C
,
Bouchoucha
YX
,
Wassef
MA
,
Gongal
PA
,
Le Men
J
,
Becker
T
, et al
Hindbrain patterning requires fine-tuning of early krox20 transcription by sprouty 4
.
Development
.
2011
;
138
:
317
26
. .
12.
Walshe
J
,
Maroon
H
,
McGonnell
IM
,
Dickson
C
,
Mason
I
.
Establishment of hindbrain segmental identity requires signaling by fgf3 and fgf8
.
Curr Biol
.
2002
;
12
:
1117
23
. .
13.
Shirasaki
R
,
Lewcock
JW
,
Lettieri
K
,
Pfaff
SL
.
Fgf as a target-derived chemoattractant for developing motor axons genetically programmed by the lim code
.
Neuron
.
2006
;
50
:
841
53
. .
14.
Esain
V
,
Postlethwait
JH
,
Charnay
P
,
Ghislain
J
.
Fgf-receptor signalling controls neural cell diversity in the zebrafish hindbrain by regulating olig2 and sox9
.
Development
.
2009
;
137
:
33
42
. .
15.
Bachler
M
,
Neubüser
A
.
Expression of members of the fgf family and their receptors during midfacial development
.
Mech Dev
.
2001
;
100
:
313
6
. .
16.
Yokoi
H
,
Shimada
A
,
Carl
M
,
Takashima
S
,
Kobayashi
D
,
Narita
T
, et al
Mutant analyses reveal different functions of fgfr1 in medaka and zebrafish despite conserved ligand–receptor relationships
.
Dev Biol
.
2007
:
326
37
.
17.
Mason
I
,
Chambers
D
,
Shamim
H
,
Walshe
J
,
Irving
C
.
Regulation and function of fgf8 in patterning of midbrain and anterior hindbrain
.
Biochem Cell Biol
.
2000
;
78
:
577
84
. .
18.
Kimmel
CB
,
Ballard
WW
,
Kimmel
SR
,
Ullmann
B
,
Schilling
TF
.
Stages of embryonic development of the zebrafish
.
Dev Dyn
.
1995
;
203
:
253
310
. .
19.
Liu
X
,
Huang
S
,
Ma
J
,
Li
C
,
Zhang
Y
,
Luo
L
.
Nf-kappab and snail1a coordinate the cell cycle with gastrulation
.
J Cell Biol
.
2009
;
184
:
805
15
. .
20.
Thummel
R
,
Bai
S
,
Sarras
MP
 Jr
,
Song
P
,
McDermott
J
,
Brewer
J
, et al
Inhibition of zebrafish fin regeneration using in vivo electroporation of morpholinos against fgfr1 and msxb
.
Dev Dyn
.
2006
;
235
:
336
46
. .
21.
Ryu
S
,
Holzschuh
J
,
Erhardt
S
,
Ettl
AK
,
Driever
W
.
Depletion of minichromosome maintenance protein 5 in the zebrafish retina causes cell-cycle defect and apoptosis
.
Proc Natl Acad Sci U S A
.
2005
;
102
:
18467
72
. .
22.
Kuo
MW
,
Lou
SW
,
Chung
BC
.
Hedgehog-PKA signaling and gnrh3 regulate the development of zebrafish gnrh3 neurons
.
PLoS One
.
2014
;
9
:
1
8
.
23.
Huang
S
,
Ma
J
,
Liu
X
,
Zhang
Y
,
Luo
L
.
Geminin is required for left-right patterning through regulating Kupffer’s vesicle formation and ciliogenesis in zebrafish
.
Biochem Biophys Res Commun
.
2011
;
410
:
164
9
. .
24.
Hwang
WY
,
Fu
Y
,
Reyon
D
,
Maeder
ML
,
Tsai
SQ
,
Sander
JD
, et al
Efficient genome editing in zebrafish using a CRISPR-Cas system
.
Nat Biotechnol
.
2013
;
31
:
227
9
. .
25.
Mali
P
,
Yang
L
,
Esvelt
KM
,
Aach
J
,
Guell
M
,
DiCarlo
JE
, et al
RNA-guided human genome engineering via Cas9
.
Science
.
2013
;
339
:
823
6
. .
26.
Huang
S
,
Ma
J
,
Liu
X
,
Zhang
Y
,
Luo
L
.
Retinoic acid signaling sequentially controls visceral and heart laterality in zebrafish
.
J Biol Chem
.
2011
;
286
:
28533
43
. .
27.
McArthur
KL
,
Fetcho
JR
.
Key features of structural and functional organization of zebrafish facial motor neurons are resilient to disruption of neuronal migration
.
Curr Biol
.
2017
;
27
(
12
):
1746.e5
56.e5
. .
28.
Bingham
S
,
Higashijima
S
,
Okamoto
H
,
Chandrasekhar
A
.
The zebrafish trilobite gene is essential for tangential migration of branchiomotor neurons
.
Dev Biol
.
2002
;
242
:
149
60
. .
29.
Hardy
CF
,
Dryga
O
,
Seematter
S
,
Pahl
PM
,
Sclafani
RA
.
mcm5/cdc46-bob1 bypasses the requirement for the S phase activator Cdc7p
.
Proc Natl Acad Sci U S A
.
1997
;
94
:
3151
5
. .
30.
Koundrioukoff
S
,
Jónsson
ZO
,
Hasan
S
,
de Jong
RN
,
van der Vliet
PC
,
Hottiger
MO
, et al
A direct interaction between proliferating cell nuclear antigen (PCNA) and Cdk2 targets PCNA-interacting proteins for phosphorylation
.
J Biol Chem
.
2000
;
275
:
22882
7
. .
31.
Boillya
B
,
Vercoutter-Edouarta
AS
,
Hondermarcka
H
,
Nurcombe
V
,
Le Bourhis
X
.
FGF signals for cell proliferation and migration through different pathways
.
Cytokine Growth Factor Rev
.
2000
;
11
:
295
302
. .
32.
Maves
L
,
Jackman
W
,
Kimmel
CB
.
Fgf3 and fgf8 mediate a rhombomere 4 signaling activity in the zebrafish hindbrain
.
Development
.
2002
;
129
:
3825
37
.
33.
Attia
L
,
Schneider
J
,
Yelin
R
,
Schultheiss
TM
.
Collective cell migration of the nephric duct requires FGF signaling
.
Dev Dyn
.
2015
;
244
:
157
67
. .
34.
Ornitz
DM
,
Itoh
N
.
The fibroblast growth factor signaling pathway
.
Wiley Interdiscip Rev Dev Biol
.
2015
;
4
:
215
66
. .
35.
Wang
X
,
Suh
C
,
Zhu
Z
,
Fan
Q
.
Minichromosome maintenance protein 5 homologue in Caenorhabditis elegans plays essential role for postembryonic development
.
Biochem Biophys Res Commun
.
2007
;
359
:
965
71
. .
36.
Yu
Z
,
Yongmei
W
,
Min
L
,
Sizhou
H
.
Expression pattern of zebrafish mcm 5 and its role in somitogenesis
.
Chin J Comp Med
.
2018
;
28
:
1671
7856
.
37.
Kok
FO
,
Shin
M
,
Ni
CW
,
Gupta
A
,
Grosse
AS
,
van Impel
A
, et al
Reverse genetic screening reveals poor correlation between morpholino-induced and mutant phenotypes in zebrafish
.
Dev Cell
.
2015
;
32
:
97
108
. .
38.
Stainier
DYR
,
Raz
E
,
Lawson
ND
,
Ekker
SC
,
Burdine
RD
,
Eisen
JS
, et al
Guidelines for morpholino use in zebrafish
.
PLoS Genet
.
2017
;
13
:
e1007000
. .
39.
Ma
Z
,
Zhu
P
,
Shi
H
,
Guo
L
,
Zhang
Q
,
Chen
Y
, et al
PTC-bearing mRNA elicits a genetic compensation response via Upf3a and compass components
.
Nature
.
2019
;
568
:
259
63
. .
40.
Bansal
A
,
Ojo
KK
,
Mu
J
,
Maly
DJ
,
Van Voorhis
WC
,
Miller
LH
.
Reduced activity of mutant calcium-dependent protein kinase 1 is compensated in plasmodium falciparum through the action of protein kinase G
.
MBio
.
2016
;
7
:
e02011
16
. .
41.
El-Brolosy
MA
,
Kontarakis
Z
,
Rossi
A
,
Kuenne
C
,
Günther
S
,
Fukuda
N
, et al
Genetic compensation triggered by mutant mRNA degradation
.
Nature
.
2019
;
568
:
193
7
. .
42.
Snyder
M
,
Huang
XY
,
Zhang
JJ
.
The minichromosome maintenance proteins 2-7 (mcm2-7) are necessary for RNA polymerase ii (pol ii)-mediated transcription
.
J Biol Chem
.
2009
;
284
:
13466
72
. .
43.
Leon
RP
,
Tecklenburg
M
,
Sclafani
RA
.
Functional conservation of beta-hairpin DNA binding domains in the mcm protein of Methanobacterium thermoautotrophicum and the mcm5 protein of saccharomyces cerevisiae
.
Genetics
.
2008
;
179
:
1757
68
. .
44.
DaFonseca
CJ
,
Shu
F
,
Zhang
JJ
.
Identification of two residues in mcm5 critical for the assembly of mcm complexes and stat1-mediated transcription activation in response to ifn-gamma
.
Proc Natl Acad Sci U S A
.
2001
;
98
:
3034
9
. .
45.
Hubbi
ME
,
Luo
W
,
Baek
JH
,
Semenza
GL
.
Mcm proteins are negative regulators of hypoxia-inducible factor 1
.
Mol Cell
.
2011
;
42
:
700
12
. .
46.
Ohtani
K
,
Iwanaga
R
,
Nakamura
M
,
Ikeda
M
,
Yabuta
N
,
Tsuruga
H
, et al
Cell growth-regulated expression of mammalian mcm5 and mcm6 genes mediated by the transcription factor e2f
.
Oncogene
.
1999
;
18
:
2299
309
.
47.
Scholpp
S
,
Groth
C
,
Lohs
C
,
Lardelli
M
,
Brand
M
.
Zebrafish fgfr1 is a member of the fgf8 synexpression group and is required for fgf8 signalling at the midbrain-hindbrain boundary
.
Dev Genes Evol
.
2004
;
214
:
285
95
. .
48.
Teraoka
H
,
Russell
C
,
Regan
J
,
Chandrasekhar
A
,
Concha
ML
,
Yokoyama
R
, et al
Hedgehog and fgf signaling pathways regulate the development of tphR-expressing serotonergic raphe neurons in zebrafish embryos
.
J Neurobiol
.
2004
;
60
:
275
88
. .
49.
Trokovic
R
,
Jukkola
T
,
Saarimaki
J
,
Peltopuro
P
,
Naserke
T
,
Weisenhorn
DM
, et al
Fgfr1-dependent boundary cells between developing mid- and hindbrain
.
Dev Biol
.
2005
;
278
:
428
39
. .
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.